LOCUS       Exported                8201 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 3'-to-5' exonuclease.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4133941
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8201)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8201)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8201
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1583..1603
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1607..1672
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1741..1862
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1752..1771)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1806..1825
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1977..2023
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2053..2189
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2234..2305
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2531..3119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2611..2630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3619..4828
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4665..4684
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4832..5860
                     /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    5887..5935
                     /note="synthetic polyadenylation signal"
     CDS             6201..7061
                     /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(6419..6438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7074..7210
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7355..7426
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      7431..7477
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7495..7543
                     /note="synthetic polyadenylation signal"
     enhancer        7624..7927
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7929..8131
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8082..8102
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      8200..8
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 caccatggaa aatgtaacag gactctttaa agattgtagt aaggtaatca ctgggttaca
       61 tcctacacag gcacctacac acctcagtgt tgacactaaa ttcaaaactg aaggtttatg
      121 tgttgacata cctggcatac ctaaggacat gacctataga agactcatct ctatgatggg
      181 ttttaaaatg aattatcaag ttaatggtta ccctaacatg tttatcaccc gcgaagaagc
      241 tataagacat gtacgtgcat ggattggctt cgatgtcgag gggtgtcatg ctactagaga
      301 agctgttggt accaatttac ctttacagct aggtttttct acaggtgtta acctagttgc
      361 tgtacctaca ggttatgttg atacacctaa taatacagat ttttccagag ttagtgctaa
      421 accaccgcct ggagatcaat ttaaacacct cataccactt atgtacaaag gacttccttg
      481 gaatgtagtg cgtataaaga ttgtacaaat gttaagtgac acacttaaaa atctctctga
      541 cagagtcgta tttgtcttat gggcacatgg ctttgagttg acatctatga agtattttgt
      601 gaaaatagga cctgagcgca cctgttgtct atgtgataga cgtgccacat gcttttccac
      661 tgcttcagac acttatgcct gttggcatca ttctattgga tttgattacg tctataatcc
      721 gtttatgatt gatgttcaac aatggggttt tacaggtaac ctacaaagca accatgatct
      781 gtattgtcaa gtccatggta atgcacatgt agctagttgt gatgcaatca tgactaggtg
      841 tctagctgtc cacgagtgct ttgttaagcg tgttgactgg actattgaat atcctataat
      901 tggtgatgaa ctgaagatta atgcggcttg tagaaaggtt caacacatgg ttgttaaagc
      961 tgcattatta gcagacaaat tcccagttct tcacgacatt ggtaacccta aagctattaa
     1021 gtgtgtacct caagctgatg tagaatggaa gttctatgat gcacagcctt gtagtgacaa
     1081 agcttataaa atagaagaat tattctattc ttatgccaca cattctgaca aattcacaga
     1141 tggtgtatgc ctattttgga attgcaatgt cgatagatat cctgctaatt ccattgtttg
     1201 tagatttgac actagagtgc tatctaacct taacttgcct ggttgtgatg gtggcagttt
     1261 gtatgtaaat aaacatgcat tccacacacc agcttttgat aaaagtgctt ttgttaattt
     1321 aaaacaatta ccatttttct attactctga cagtccatgt gagtctcatg gaaaacaagt
     1381 agtgtcagat atagattatg taccactaaa gtctgctacg tgtataacac gttgcaattt
     1441 aggtggtgct gtctgtagac atcatgctaa tgagtacaga ttgtatctcg atgcttataa
     1501 catgatgatc tcagctggct ttagcttgtg ggtttacaaa caatttgata cttataacct
     1561 ctggaacact tttacaagac ttgaaaacct ttactttcag tcgatggatt ataaggacca
     1621 cgatggagac tacaaagatc atgacattga ttacaaggac gacgatgaca aataaggatc
     1681 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
     1741 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1801 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1861 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1921 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
     1981 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
     2041 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     2101 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     2161 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
     2221 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
     2281 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
     2341 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
     2401 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
     2461 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2521 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2581 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2641 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2701 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2761 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2821 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2881 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2941 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3001 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3061 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3121 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3181 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3241 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3301 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     3361 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     3421 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     3481 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     3541 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     3601 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     3661 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     3721 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     3781 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     3841 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     3901 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     3961 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     4021 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     4081 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     4141 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     4201 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     4261 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     4321 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     4381 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     4441 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     4501 cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     4561 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     4621 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     4681 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     4741 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     4801 ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac
     4861 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     4921 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     4981 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     5041 cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta
     5101 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     5161 cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca
     5221 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     5281 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     5341 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     5401 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     5461 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     5521 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     5581 cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat
     5641 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     5701 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     5761 cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     5821 aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc
     5881 ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa
     5941 cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     6001 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     6061 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     6121 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     6181 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     6241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6301 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6361 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     6421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6481 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     6541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6601 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     6781 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     6841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7021 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     7081 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     7141 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     7201 ttatccttaa gatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct
     7261 tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc
     7321 agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt
     7381 atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc
     7441 ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca
     7501 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga
     7561 gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat atatggagtt
     7621 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     7681 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     7741 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     7801 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     7861 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     7921 taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     7981 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     8041 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     8101 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt
     8161 gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
