LOCUS       sequence-202318-        4717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid of bdSENP1 protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4717)
  AUTHORS   Frey S, Gorlich D
  TITLE     A new set of highly efficient, tag-cleaving proteases for purifying 
            recombinant proteins.
  JOURNAL   J Chromatogr A. 2014 Apr 11;1337:95-105. doi: 
            10.1016/j.chroma.2014.02.029. Epub 2014 Feb 19.
  PUBMED    24636565
REFERENCE   2  (bases 1 to 4717)
  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: "J 
            Chromatogr A."; date: "2014-04-11"; volume: "1337:95-105. doi"; 
            pages: " 10.1016/j.chroma.2014.02.029. Epub 2014 Feb 19"
COMMENT     Sequence Label: pSF1389
FEATURES             Location/Qualifiers
     source          1..4717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     35..53
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(77..91,92..97,98..114,115..121)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    97..113
                     /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)."
     protein_bind    129..145
                     /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             164..175
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             260..280
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      1029..1123
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1171..1190)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1182..1997
                     /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             1182..1997
                     /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     complement(1255..1274)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2216..2302
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2395..3477)
                     /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     3439..3458
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3478..3555)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(3639..3661)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3747..3887
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3802..3821
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3902..3919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4073..4661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4153..4172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaagggatt ttggtcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
       61 tcttcacctc gagaaatcat aaaaaattta tttgctttgt gagcggataa caattataat
      121 agattcaatt gtgagcggat aacaatttca cacagaattc attaaagagg agaaattaac
      181 catgagcaag catcaccatc attcaggcca tcaccatacc ggacaccacc atcattcagg
      241 cagtcatcac cataccggcg agaacctgta ttttcagggt accaaggaag aagtgccgga
      301 accgtttgtg ccgttgaccg acgaagacga agataacgtt cgtcacgcac tgggtggccg
      361 caaacgttcg gaaaccctgt ccgtgcacga agcgtcaaat attgtgatca cgcgcgagat
      421 cctgcagtgt ctgaacgaca aagaatggct gaatgatgag gttatcaacc tgtacctgga
      481 actgctgaaa gagcgtgaac tgcgtgaacc gaacaaattt ctgaaatgtc atttcttcaa
      541 tacgtttttc tacaaaaagc tgattaacgg cggctatgat tacaaaagcg ttcgtcgctg
      601 gactacgaaa cgcaaactgg gttacaacct gattgattgc gacaaaatct tcgtgccgat
      661 tcacaaagat gttcattggt gcctggctgt aattaacatc aaagaaaaga aattccagta
      721 tctcgatagc ctgggctata tggatatgaa agcgctgcgc attctggcta aatatctcgt
      781 ggacgaagta aaagacaaat ccggtaagca gatcgacgtt cacgcgtgga aacaggaggg
      841 tgttcagaac ctgccgctgc aggaaaacgg ttgggattgc ggtatgttca tgctgaagta
      901 tatcgatttc tattctcgtg acatggaact ggtctttggc cagaaacaca tgtcgtactt
      961 tcgtcgtcgt accgctaaag aaatcctgga tctgaaggcc ggttagtaag cttaattagc
     1021 tgagcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     1081 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagct agccatgaaa
     1141 ataaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     1201 aaacgtcttg ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     1261 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc
     1321 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     1381 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     1441 ttatccgtac tcctgatgat gcttggttac tcacgactgc gatccccggc aaaacagcat
     1501 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     1561 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     1621 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     1681 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcac aaacttttgc
     1741 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     1801 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     1861 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     1921 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     1981 tcgatgagtt tttctaagaa ttaattcatg ggcaaatatt atacgcaagg cgacaaggtg
     2041 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     2101 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa
     2161 ggcagttatt ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat
     2221 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     2281 cgctctcctg agtaggacaa atccgccctc tagattacgt gcagtcgatg ataagctgtc
     2341 aaacatgaga attgtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     2401 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2461 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     2521 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     2581 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     2641 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     2701 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     2761 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     2821 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     2881 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     2941 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     3001 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     3061 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     3121 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     3181 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     3241 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     3301 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     3361 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     3421 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     3481 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca
     3541 ccattcgatg gtgtcggaat ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     3601 tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     3661 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     3721 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     3781 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     3841 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     3901 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3961 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4021 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4081 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4141 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4201 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4261 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4321 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4381 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4441 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4501 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4561 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4621 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4681 ctacggggtc tgacgctcag tggaacgaaa actcacg
//
