LOCUS       sequence-380198-        5960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses high-efficiency TEV protease in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5960)
  AUTHORS   Lau K, Bouchri B, Pojer F
  TITLE     Purification of 10xHis-SuperTEV
  JOURNAL   protocols.io, doi: 10.17504/protocols.io.yxmvm2zxng3p/v1
REFERENCE   2  (bases 1 to 5960)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "protocols.io, doi: 10.17504/protocols.io.yxmvm2zxng3p/v1"
COMMENT     Sequence Label: pET29b-10xHis-Super TEV
FEATURES             Location/Qualifiers
     source          1..5960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..34
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        15..33
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    34..58
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             73..95
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             109..138
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             835..852
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(905..923)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      919..966
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1003..1458
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1090..1109)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1300..1321
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1550..2365)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1550..2365)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2273..2292
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2357..2376
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2487..3075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2976..2995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3229..3246
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3261..3400
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3326..3345)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3486..3508
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3502..3693)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4502..5584)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5546..5565
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5585..5662)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5868..5887
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
       61 tctagaaata attttgttta actttaagaa ggagatatac atatgaaaca tcaccatcat
      121 caccaccatc atcatcatgg atccggagaa agcttgttta agggaccacg tgattacaac
      181 ccgatatcga gcagcatttg tcatttgacg aatgaatctg atgggcacac aacatcgttg
      241 tatggtattg gatttggtcc cttcatcatt acaaacaagc acttgtttcg tcgcaataat
      301 ggaacactgg tagtccaatc actacatggt gtattcaagg tcaaggacac cgcgactttg
      361 caacaacacc tcgttgatgg gagggacatg atgattattc gcatgcctaa ggatttccca
      421 ccatttcctc aaaagctgaa atttagagag ccacaaaggg aagagcgcat atgtcttgtg
      481 acaaccaact tccaagctaa gagcatgtct agcatggtgt cagacactag ttgcacattc
      541 ccttcagggg atggtacatt ctggaagcat tggattcaaa ccaaggatgg gcagtgtggc
      601 aatccattag tatcaactag agatgggttc attgttggta tacactcagc atcgaatttc
      661 accaacacaa acaattattt cgcaagcgtg ccgaaaaact tcatggaatt gttgacaaat
      721 caggaggcgc agcagtgggt tagtggttgg cgattaaacg ctgactcagt attgtggggg
      781 ggccataaag ttttcatggt caaacctgaa gaacctggtt cttaatagct cgagcaccac
      841 caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
      901 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
      961 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
     1021 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     1081 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     1141 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     1201 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     1261 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     1321 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     1381 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     1441 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1501 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
     1561 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     1621 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     1681 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     1741 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     1801 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     1861 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     1921 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     1981 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     2041 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     2101 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     2161 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2221 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     2281 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
     2341 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     2401 gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2461 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2521 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2581 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     2641 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2701 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2761 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2821 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2881 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2941 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3001 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3061 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3121 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3181 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3241 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3301 tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     3361 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     3421 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3481 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     3541 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     3601 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     3661 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     3721 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     3781 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     3841 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     3901 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     3961 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     4021 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     4081 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     4141 cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa
     4201 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
     4261 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
     4321 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
     4381 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     4441 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     4501 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4561 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     4621 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     4681 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     4741 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca
     4801 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     4861 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     4921 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     4981 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     5041 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     5101 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     5161 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     5221 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     5281 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     5341 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     5401 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     5461 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     5521 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     5581 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     5641 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     5701 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     5761 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     5821 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     5881 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     5941 tagaggatcg agatcgatct
//
