LOCUS       Exported                7167 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4134148
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7167)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7167)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7167
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        200..402
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     353..373
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      471..480
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1014..1034
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1038..1103
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1172..1293
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1183..1202)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1237..1256
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1408..1454
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1484..1620
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1665..1736
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1962..2550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2042..2061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3050..4259
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4096..4115
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4263..5291
                     /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    5318..5366
                     /note="synthetic polyadenylation signal"
     CDS             5632..6492
                     /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(5850..5869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6505..6641
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6793..6864
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6869..6915
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6933..6981
                     /note="synthetic polyadenylation signal"
     enhancer        7062..198
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
       61 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      121 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      181 gtcatcgcta ttaccatgct gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      241 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      301 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      361 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
      421 atcagatctt tgtcgatcct accatccact cgacacaccc gccagcggcc gccaccatgg
      481 agagccttgt ccctggtttc aacgagaaaa cacacgtcca actcagtttg cctgttttac
      541 aggttcgcga cgtgctcgta cgtggctttg gagactccgt ggaggaggtc ttatcagagg
      601 cacgtcaaca tcttaaagat ggcacttgtg gcttagtaga agttgaaaaa ggcgttttgc
      661 ctcaacttga acagccctat gtgttcatca aacgttcgga tgctcgaact gcacctcatg
      721 gtcatgttat ggttgagctg gtagcagaac tcgaaggcat tcagtacggt cgtagtggtg
      781 agacacttgg tgtccttgtc cctcatgtgg gcgaaatacc agtggcttac cgcaaggttc
      841 ttcttcgtaa gaacggtaat aaaggagctg gtggccatag ttacggcgcc gatctaaagt
      901 catttgactt aggcgacgag cttggcactg atccttatga agattttcaa gaaaactgga
      961 acactaaaca tagcagtggt gttacccgtg aactcatgcg tgagcttaac ggagaaaacc
     1021 tttactttca gtcgatggat tataaggacc acgatggaga ctacaaagat catgacattg
     1081 attacaagga cgacgatgac aaataaggat ccgcaggcct ctgctagctt gactgactga
     1141 gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc
     1201 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1261 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1321 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     1381 ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct
     1441 tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa
     1501 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     1561 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     1621 ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc
     1681 ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct
     1741 cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga
     1801 tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct
     1861 tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag
     1921 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1981 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2341 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2401 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2461 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2521 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2581 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2641 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2701 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2761 tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg
     2821 gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg
     2881 caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat
     2941 tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc
     3001 aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc
     3061 cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac
     3121 gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct
     3181 cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg
     3241 gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc
     3301 ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg
     3361 acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt
     3421 ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt
     3481 cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag
     3541 tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc
     3601 ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac
     3661 aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg
     3721 aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac gtgaagtttg
     3781 tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg
     3841 ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg
     3901 ttctgttggc ttataatgca gggtggggcc acctgccggt aggtgtgcgg taggcttttc
     3961 tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg
     4021 acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct
     4081 atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt
     4141 tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc
     4201 agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagaca
     4261 acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     4321 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     4381 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     4441 aagatcgtta tgtttatcgg cactttgcat ccgccgcgct cccgattccg gaagtgcttg
     4501 acattgggga gttcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     4561 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcaa
     4621 tggatgccat cgctgccgcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     4681 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     4741 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     4801 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     4861 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     4921 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     4981 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     5041 cgccccggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     5101 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     5161 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgccgcc gtctggaccg
     5221 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     5281 caaaggaata agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag
     5341 atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg
     5401 atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat
     5461 ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag
     5521 cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc
     5581 ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt
     5641 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5701 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt
     5761 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc
     5821 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5881 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
     5941 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6001 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg
     6061 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6121 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6181 atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6241 cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6301 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6361 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6421 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6481 aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg
     6541 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     6601 cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac
     6661 tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc
     6721 atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat
     6781 cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca
     6841 taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc
     6901 caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct
     6961 tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca
     7021 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     7081 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     7141 gttcccatag taacgccaat agggact
//
