LOCUS       sequence-430273-        4717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of SENP_EuB protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4717)
  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 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"; doi: 
            "10.1083/jcb.201812091"; journalName: "J Cell Biol"; date: 
            "2019-06-3- 3"; volume: "218"; issue: "6"; pages: "2006-2020"
COMMENT     Sequence Label: pAV0286
FEATURES             Location/Qualifiers
     source          1..4717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..15,16..21,22..38,39..45)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    21..37
                     /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    53..69
                     /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             88..99
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             184..204
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      953..1047
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1095..1114)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1106..1921
                     /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             1106..1921
                     /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(1179..1198)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2140..2226
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2319..3401)
                     /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     3363..3382
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3402..3479)
                     /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(3563..3585)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3671..3811
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3726..3745
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3826..3843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3997..4585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4077..4096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4676..4694
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
       61 ggataacaat ttcacacaga attcattaaa gaggagaaat taaccatgag caagcatcac
      121 catcattcag gccatcacca taccggacac caccatcatt caggcagtca tcaccatacc
      181 ggcgagaacc tgtattttca gggtaccaag gaagaagtgc cggaaccgtt tgtgccgttg
      241 accgacgaag acgaagataa cgttcgtcac gcactgggtg gccgcaaacg ttccgaaacc
      301 ctgtccgtgc acgaagcgtc atctattgtg atcacgcgcg agatcctgca gtgtctgaac
      361 gacaaagaat ggctgaatga tgaggttatc aacctgtacc tggaactgct gaaagagcgt
      421 gaactgcgtg aaccgaacaa atttctgaaa tgtcatttct tcaatacgtt tttctacaaa
      481 aagctgatta acggcggcta tgattacaaa agcgttcgtg agtggactac gaagcgcaaa
      541 ctgggttaca acctgattga ttgcgacaaa atcttcgtgc cgattcacaa agatgttcat
      601 tggtgcctgg ctgtaattaa catcaaagaa aagaaattcc agtatctcga tagcctgggc
      661 tatatggata tgaaagcgct gcgcattctg gctaaatatc tcgtggacga agtaaaagac
      721 aaatccggta agcagatcga cgttcacgcg tggaaacagg agggtgttca gaacctgccg
      781 ctgcaggaaa acggttggga ttgcggtatg ttcatgctga agtatatcga tttctattct
      841 cgtgacatgg aactggtctt tggccagaaa cacatgtcgt actttcgtcg tcgtaccgct
      901 aaagaaatcc tggatctgaa ggccggttag taagcttaat tagctgagct tggactcctg
      961 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     1021 cgccgggcgt tttttattgg tgagaatcca agctagccat gaaaataaac tgtctgctta
     1081 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctctag
     1141 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     1201 tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt
     1261 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     1321 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     1381 tgatgcttgg ttactcacga ctgcgatccc cggcaaaaca gcattccagg tattagaaga
     1441 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     1501 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     1561 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     1621 ctggcctgtt gaacaagtct ggaaagaaat gcacaaactt ttgccattct caccggattc
     1681 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     1741 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     1801 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     1861 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     1921 agaattaatt catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     1981 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     2041 aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc
     2101 cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag
     2161 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     2221 acaaatccgc cctctagatt acgtgcagtc gatgataagc tgtcaaacat gagaattgtg
     2281 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2341 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2401 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     2461 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     2521 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     2581 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     2641 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     2701 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     2761 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     2821 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     2881 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     2941 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     3001 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     3061 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     3121 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     3181 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     3241 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     3301 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     3361 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     3421 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcaccattc gatggtgtcg
     3481 gaatttcggg cagcgttggg tcctggccac gggtgcgcat gatctagagc tgcctcgcgc
     3541 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     3601 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     3661 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     3721 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     3781 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     3841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4621 tcagtggaac gaaaactcac gttaagggat tttggtcatg acattaacct ataaaaatag
     4681 gcgtatcacg aggccctttc gtcttcacct cgagaaa
//
