LOCUS       sequence-295681-        7136 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7136)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7136)
  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-m4134386
FEATURES             Location/Qualifiers
     source          1..7136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             545..565
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             566..607
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    676..797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(687..706)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     741..760
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      912..958
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      988..1124
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1169..1240
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1466..2054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1546..1565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2554..3763
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3600..3619
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3767..4795
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4822..4870
                     /note="synthetic polyadenylation signal"
     CDS             join(5136..5204,5205..5996)
                     /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(5354..5373)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6009..6145
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6290..6361
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6366..6412)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6430..6478
                     /note="synthetic polyadenylation signal"
     enhancer        6559..6862
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6864..7066
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7017..7037
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7135..7136,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggag tctctcgttc ccggctttaa tgaaaaaact catgtccagt tatccttgcc
       61 agtgcttcaa gtaagggatg tgctggtgcg gggtttcgga gacagtgtag aggaagtcct
      121 gtcagaggct agacagcacc tgaaggatgg gacctgtggc ttagtggaag ttgagaaggg
      181 agtcctcccg cagctggaac aaccttatgt gtttataaaa cgcagcgacg cgcgaacagc
      241 acctcacggg catgtaatgg tggagcttgt cgccgagttg gagggcatcc agtacggaag
      301 atcgggtgaa acgctgggag ttctagtgcc ccacgttggc gaaattccgg tggcctaccg
      361 taaggttttg ctcaggaaaa acggaaataa gggggctggt ggccatagct atggggcaga
      421 cctgaagtct ttcgatctgg gcgacgagct gggcaccgac ccatatgagg atttccagga
      481 aaactggaac acaaaacaca gctccggggt aactcgcgag ctaatgcgag aattgaatgg
      541 aggggaaaac ctttactttc agtcggggaa acctataccc aatccacttc tcggcttaga
      601 ttctacctaa ggatccgcag gcctctgcta gcttgactga ctgagataca gcgtaccttc
      661 agctcacaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
      721 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
      781 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
      841 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc
      901 tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg
      961 cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg
     1021 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     1081 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct
     1141 gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact
     1201 aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg
     1261 ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt
     1321 tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt
     1381 ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc
     1441 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1501 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1561 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1621 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1681 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1741 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1801 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1861 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1921 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1981 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2041 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2101 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2161 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2221 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2281 ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct
     2341 ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg
     2401 ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat
     2461 atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg
     2521 agatccgagg aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc
     2581 ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg
     2641 tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt
     2701 agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact
     2761 ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc
     2821 ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc
     2881 acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga
     2941 tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg
     3001 ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca
     3061 aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct
     3121 tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg
     3181 tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg cgggaaagct cttattcggg
     3241 tgagatgggc tgggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac
     3301 tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg
     3361 tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa
     3421 tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca
     3481 gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga
     3541 gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct
     3601 gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta
     3661 ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agacttgtaa
     3721 attgtccgct aaattctggc cgtttttggc ttttttgtta gacaacatgg gtaaaaagcc
     3781 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     3841 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     3901 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     3961 tcggcacttt gcatccgccg cgctcccgat tccggaagtg cttgacattg gggagttcag
     4021 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     4081 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gcaatggatg ccatcgctgc
     4141 cgccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     4201 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     4261 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     4321 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     4381 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     4441 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     4501 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgcccc ggctccgggc
     4561 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     4621 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     4681 cgggcgtaca caaatcgccc gcagaagcgc cgccgtctgg accgatggct gtgtagaagt
     4741 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aataagctag
     4801 tatgtaagcc tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt
     4861 tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag
     4921 tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag
     4981 gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta
     5041 aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca
     5101 tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg
     5161 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5221 tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc
     5281 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca
     5341 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5401 tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa
     5461 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5521 ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt
     5581 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5641 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc
     5701 gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga
     5761 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5821 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5881 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5941 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga
     6001 ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     6061 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     6121 ctccttacca gaaatttatc cttaagatcg tttaaactcg actctggctc tatcgaatct
     6181 ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc
     6241 tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc
     6301 attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct
     6361 caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt
     6421 tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat
     6481 ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag
     6541 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     6601 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     6661 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     6721 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     6781 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     6841 attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata
     6901 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     6961 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     7021 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     7081 tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgc
//
