LOCUS       sequence-306146-        4723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of SENP_EuH protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4723)
  AUTHORS   Vera Rodriguez A, Frey S, Gorlich D
  TITLE     Engineered SUMO/protease system identifies Pdr6 as a bidirectional 
            nuclear transport receptor.
  JOURNAL   J Cell Biol. 2019 Jun 3;218(6):2006-2020. doi: 
            10.1083/jcb.201812091. Epub 2019 Apr 25.
  PUBMED    31023724
REFERENCE   2  (bases 1 to 4723)
  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"; doi: 
            "10.1083/jcb.201812091"; journalName: "J Cell Biol"; date: 
            "2019-06-3- 3"; volume: "218"; issue: "6"; pages: "2006-2020"
COMMENT     Sequence Label: pAV0679
FEATURES             Location/Qualifiers
     source          1..4723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..54)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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    62..78
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             193..213
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      968..1062
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1110..1129)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1121..1936
                     /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             1121..1936
                     /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(1194..1213)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2155..2241
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2334..3416)
                     /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     3378..3397
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3417..3494)
                     /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(3578..3600)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3686..3826
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3741..3760
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3841..3858)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4012..4600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4092..4111)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4691..4709
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aaccatgagc
      121 aagcatcacc atcattcagg ccatcaccat accggacacc accatcattc aggcagtcat
      181 caccataccg gcgagaacct gtattttcaa gggacgtccg gtaccaagga agaagtgccg
      241 gaaccgtttg tgccgttgac cgacgaagac gaagataacg ttcgtcacgc actgggtggc
      301 cgcaaaccgt cggaaaccct gtccgtgcac gaagcgtcag ctattgtgat cacgcgcgag
      361 atcctgcagt gtctgaacga caaagaatgg ctgaatgatg aggttatcaa cctgtacctg
      421 gaactgctga aagagcgtga actgcgtgaa ccgaacaaat ttctgaaatg tcatttcttc
      481 aatacgtttt tctacaaaaa gctgattaac ggctatgatt acaaaagcgt tcgtgagtgg
      541 actacgatgc gcaaactggg ttacaacctg attgattgcg acaaaatctt cgtgccgatt
      601 cacaaagatg ttcattggtg cctggctgta attaacatca aagaaaagaa attccagtat
      661 ctcgatagcc tgggctatat ggatatgaaa gcgctgcgca ttctggctaa atatctcgtg
      721 gacgaagtaa aagacaaatc cggtaagcag atcgacgttc acgcgtggaa acaggagggt
      781 gttcagaacc tgccgctgca ggaaaacggt tgggattgcg gtatgttcat gctgaagtat
      841 atcgatttct attctcgtga catggaactg gtctttggcc agaaacacat gtcgtacttt
      901 cgtcgtcgta ccgctaaaga aatcctggat ctgaaggccg gttagtaagc ttaattagct
      961 gagcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
     1021 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta gccatgaaaa
     1081 taaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     1141 aacgtcttgc tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     1201 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
     1261 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     1321 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     1381 tatccgtact cctgatgatg cttggttact cacgactgcg atccccggca aaacagcatt
     1441 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     1501 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     1561 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     1621 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcaca aacttttgcc
     1681 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     1741 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     1801 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     1861 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     1921 cgatgagttt ttctaagaat taattcatgg gcaaatatta tacgcaaggc gacaaggtgc
     1981 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     2041 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
     2101 gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag gcatcaaata
     2161 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     2221 gctctcctga gtaggacaaa tccgccctct agattacgtg cagtcgatga taagctgtca
     2281 aacatgagaa ttgtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     2341 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2401 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     2461 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     2521 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     2581 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     2641 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     2701 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     2761 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     2821 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     2881 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     2941 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     3001 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     3061 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     3121 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     3181 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     3241 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     3301 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     3361 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     3421 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac
     3481 cattcgatgg tgtcggaatt tcgggcagcg ttgggtcctg gccacgggtg cgcatgatct
     3541 agagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     3601 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     3661 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     3721 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     3781 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
     3841 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3901 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3961 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4021 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4081 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4141 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4201 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4261 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4321 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4381 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4441 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4501 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4561 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4621 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgacatt
     4681 aacctataaa aataggcgta tcacgaggcc ctttcgtctt cac
//
