LOCUS       Exported                7109 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4134220
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7109)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7109)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7109
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             542..562
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             563..580
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    649..770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(660..679)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     714..733
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      885..931
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      961..1097
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1142..1213
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1439..2027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1519..1538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2527..3736
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3573..3592
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3740..4768
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4795..4843
                     /note="synthetic polyadenylation signal"
     CDS             5109..5969
                     /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(5327..5346)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5982..6118
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6263..6334
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6339..6385
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6403..6451
                     /note="synthetic polyadenylation signal"
     enhancer        6532..6835
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6837..7039
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6990..7010
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      7108..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggag agccttgtcc ctggtttcaa cgagaaaaca cacgtccaac tcagtttgcc
       61 tgttttacag gttcgcgacg tgctcgtacg tggctttgga gactccgtgg aggaggtctt
      121 atcagaggca cgtcaacatc ttaaagatgg cacttgtggc ttagtagaag ttgaaaaagg
      181 cgttttgcct caacttgaac agccctatgt gttcatcaaa cgttcggatg ctcgaactgc
      241 acctcatggt catgttatgg ttgagctggt agcagaactc gaaggcattc agtacggtcg
      301 tagtggtgag acacttggtg tccttgtccc tcatgtgggc gaaataccag tggcttaccg
      361 caaggttctt cttcgtaaga acggtaataa aggagctggt ggccatagtt acggcgccga
      421 tctaaagtca tttgacttag gcgacgagct tggcactgat ccttatgaag attttcaaga
      481 aaactggaac actaaacata gcagtggtgt tacccgtgaa ctcatgcgtg agcttaacgg
      541 agaaaacctt tactttcagt cgcaccatca ccatcaccat taaggatccg caggcctctg
      601 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
      661 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      721 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
      781 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
      841 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
      901 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
      961 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     1021 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     1081 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     1141 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     1201 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     1261 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     1321 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     1381 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1441 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1501 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1561 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1621 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1681 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1741 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1801 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1861 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1921 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1981 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2041 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2101 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2161 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2221 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     2281 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     2341 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     2401 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     2461 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     2521 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     2581 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     2641 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     2701 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     2761 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     2821 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     2881 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     2941 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     3001 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     3061 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     3121 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     3181 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     3241 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     3301 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     3361 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     3421 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     3481 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     3541 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     3601 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     3661 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     3721 ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     3781 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     3841 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     3901 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc
     3961 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     4021 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     4081 gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt
     4141 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     4201 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     4261 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     4321 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     4381 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     4441 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     4501 tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca
     4561 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     4621 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     4681 cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     4741 cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa
     4801 atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc
     4861 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     4921 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     4981 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     5041 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     5101 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5161 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5221 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5281 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5341 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5401 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5461 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5521 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5581 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5641 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     5701 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     5761 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5821 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5881 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5941 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     6001 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     6061 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     6121 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     6181 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     6241 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     6301 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     6361 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     6421 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     6481 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     6541 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     6601 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     6661 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     6721 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     6781 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat
     6841 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     6901 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     6961 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     7021 ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt cgatcctacc
     7081 atccactcga cacacccgcc agcggccgc
//
