LOCUS       Exported                6204 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp2.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4134416
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6204)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6204)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6204
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2500..2520
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2521..2598
                     /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"
     polyA_signal    2651..2772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2662..2681)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2716..2735
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2887..2933
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2963..3099
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3144..3215
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3441..4029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3521..3540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4775..5635
                     /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(4993..5012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5648..5784
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5936..6007
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6012..6058
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6076..6124
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggccta cacaagatat
      601 gtggataaca atttttgtgg gccggacgga taccctttag aatgcatcaa agatttgctt
      661 gcaagggcgg gcaaggcttc gtgcactctg tccgagcagc tggacttcat tgataccaag
      721 cgcggtgttt attgttgccg agagcatgaa cacgagatag cctggtatac ggaacggtca
      781 gagaaaagtt acgaactcca aaccccattc gagatcaagc tagctaaaaa gtttgacact
      841 ttcaacggcg aatgtcccaa ttttgtcttt cccctgaact ctattatcaa aacaatacag
      901 cctcgggtgg agaagaagaa attagacgga ttcatggggc gtattaggag cgtataccca
      961 gtggccagcc cgaatgaatg caaccagatg tgtctttcca cactgatgaa gtgcgatcac
     1021 tgtggcgaga cttcttggca aaccggtgac ttcgtaaaag cgacatgcga gttttgtggt
     1081 accgagaatc tgactaagga gggagctacc acatgcggct atttgccaca gaatgcagtg
     1141 gtcaagatct actgtcctgc atgccataac tctgaagtcg gccccgagca ctcgctcgcc
     1201 gaatatcaca atgagagcgg gctaaaaact atactgagga aaggaggtcg caccatcgcc
     1261 tttgggggct gcgttttctc ctacgtgggg tgtcataaca aatgtgccta ttgggttccc
     1321 agagcctccg ctaacattgg gtgcaatcat acgggagtgg tcggggaagg ctcagaaggc
     1381 ttgaacgata acctccttga aattctgcaa aaggagaagg tgaatatcaa tatcgttggc
     1441 gacttcaagt taaatgagga aatcgcgatt attctggcaa gtttttctgc ctcaactagc
     1501 gcctttgtag agacagtgaa aggactcgat tataaagctt tcaaacagat agtcgagagt
     1561 tgtggtaact tcaaggttac aaagggcaag gctaaaaagg gtgcttggaa cattggagag
     1621 cagaagtcaa tcttgagtcc actgtacgcg tttgcatctg aggccgcccg tgtggtcagg
     1681 agcatattca gtcgaaccct agaaaccgca cagaatagcg tgcgggtatt acagaaggcg
     1741 gctattacta ttctcgacgg tatctcgcag tattctctga gactgatcga cgccatgatg
     1801 tttacaagcg atctggcaac gaataactta gtggtcatgg cgtacataac cggaggggtc
     1861 gtgcaactta cctcccagtg gctaacgaac atctttggga cggtttacga gaagctaaaa
     1921 cctgtgctgg actggctcga agagaaattc aaagaaggcg tggaattttt gcgtgacggg
     1981 tgggagatcg taaagttcat atcaacatgt gcttgcgaaa tcgttggcgg acaaatcgta
     2041 acttgcgcaa aagagattaa agaatcggtg cagaccttct ttaaacttgt taacaaattc
     2101 ctggctttgt gtgccgattc tattatcatc gggggagcaa agcttaaagc cctaaattta
     2161 ggcgagacat ttgtgacgca cagtaagggc ctgtatagaa agtgcgtgaa gagtcgcgaa
     2221 gaaaccggac tgctcatgcc cttaaaagcg ccaaaagaaa taattttcct ggaaggggag
     2281 acattgccca ctgaagtgct cactgaggaa gtagttctga aaactgggga tctgcaacct
     2341 ttagagcagc cgacctccga ggccgtcgag gcacctctcg tgggaacccc ggtctgtatt
     2401 aatgggttga tgctgctcga gattaaggat accgaaaaat actgcgcact agcccccaac
     2461 atgatggtga ctaacaacac atttaccctc aagggagggg aaaaccttta ctttcagtcg
     2521 aagaggcgat ggaaaaagaa ttttatcgcg gtgagtgcag ccaacagatt caaaaagata
     2581 tcctcgtctg gtgccctcta agctagcttg actgactgag atacagcgta ccttcagctc
     2641 acagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2701 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2761 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     2821 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg
     2881 gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct
     2941 cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     3001 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     3061 cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg
     3121 cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg
     3181 tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc
     3241 gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac
     3301 tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg
     3361 attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3841 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4261 gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt
     4321 ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg
     4381 cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg
     4441 agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc
     4501 cgaggaatat cgaaatcggg gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt
     4561 ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt gggacccagg
     4621 aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa
     4681 acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt ttgcaaacat
     4741 ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt tccgtgtcgc
     4801 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4861 gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct
     4921 caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac
     4981 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5041 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
     5101 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5161 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
     5221 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5281 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg
     5341 taaactatta actggcgaac tacttactct agcttcccgg caacagttga tagactggat
     5401 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5461 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5521 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5581 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat
     5641 tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     5701 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     5761 tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc tggctctatc
     5821 gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat
     5881 cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct cccgacatct
     5941 tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc agcttcagct
     6001 acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctatc
     6061 aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt
     6121 tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg ggtcattagt
     6181 tcatagccca tatatggagt tccg
//
