LOCUS       sequence-138319-        5973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5973)
  AUTHORS   Mikolajczyk J, Drag M, Bekes M, Cao JT, Ronai Z, Salvesen GS
  TITLE     Small ubiquitin-related modifier (SUMO)-specific proteases: 
            profiling the specificities and activities of human SENPs.
  JOURNAL   J Biol Chem. 2007 Sep 7. 282(36):26217-24.
  PUBMED    17591783
REFERENCE   2  (bases 1 to 5973)
  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 Biol 
            Chem. 2007 Sep 7. 282(36):26217-24."
COMMENT     Sequence Label: pET28a-SENP1 (catalytic domain)
FEATURES             Location/Qualifiers
     source          1..5973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(88..107)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        313..390
                     /gene="lacI"
                     /label=lacI promoter
     CDS             391..1473
                     /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     complement(410..429)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2282..2473
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2467..2489)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2575..2714
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2630..2649
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2729..2746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2900..3488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2980..2999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3599..3618)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3610..4425
                     /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             3610..4425
                     /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(3683..3702)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(4517..4972)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4654..4675)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4866..4885
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(5009..5056)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     5052..5070
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(5123..5140)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5834..5851)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(5861..5878)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             5897..5919
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5934..5958
                     /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)."
     primer_bind     complement(join(5958..5973,1..4))
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(join(5959..5973,1..4))
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 attaatttcg cgggatcgag atctcgatcc tctacgccgg acgcatcgtg gccggcatca
       61 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
      121 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
      181 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc
      241 tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc
      301 gtcgagatcc cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc
      361 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca
      421 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
      481 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
      541 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
      601 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
      661 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
      721 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
      781 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
      841 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
      901 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
      961 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
     1021 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
     1081 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
     1141 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta
     1201 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
     1261 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
     1321 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
     1381 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     1441 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct
     1501 cactcattag gcaccgggat ctcgaccgat gcccttgaga gccttcaacc cagtcagctc
     1561 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     1621 gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg
     1681 ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc
     1741 tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc
     1801 cggcatggcg gccccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg
     1861 ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag
     1921 cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg
     1981 tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg
     2041 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg
     2101 gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc
     2161 ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct
     2221 ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc
     2281 agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt
     2341 aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc
     2401 gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg
     2461 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2521 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     2581 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
     2641 cagattgtac tgagagtgca ccattgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2701 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2761 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2821 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2881 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2941 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3001 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3061 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3121 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3181 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3241 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3301 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3361 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3421 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3481 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3541 cacgttaagg gattttggtc atgaacaata aaactgtctg cttacataaa cagtaataca
     3601 aggggtgtta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg attaaattcc
     3661 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     3721 gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     3781 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     3841 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     3901 accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt
     3961 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     4021 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     4081 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     4141 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     4201 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     4261 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     4321 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     4381 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagaatt aattcatgag
     4441 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4501 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4561 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4621 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4681 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4741 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4801 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4861 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4921 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     4981 ccaatccgga tatagttcct cctttcagca aaaaacccct caagacccgt ttagaggccc
     5041 caaggggtta tgctagttat tgctcagcgg tggcagcagc caactcagct tcctttcggg
     5101 ctttgttagc agccggatct cagtggtggt ggtggtggtg ctcgagtcac aagagttttc
     5161 ggtggaggat ctcccagacc atccgcttcc ggaagtatgg catgtgttgc tgtgtgaagt
     5221 tgattggtct gtctttggta atacagtcag catatttgca ggcaaacatc ccacagtcac
     5281 ttccattcat ctgctgagga atctcctggc ttttcttgct gaaaagctgc cagccattgg
     5341 tgtcaaactc tttccttttc ttgtcaatgc tttcttgctt taggtattgc aagagtattc
     5401 tgcaggcttc attgtttatc ccacccatgg agtcgtaata ggtaatattc ttctttctaa
     5461 agtccacaac agctagacac cagtgtactc ccaggtgaat gggcaccaaa agaatgtcaa
     5521 cagaaaatac atctactttc tttgtccaac gtttcactgc ctgataacca gccgttttta
     5581 atttagtgaa gaaaaaggta ttaaatgcat gcacacttgg caagcccttc tctttacttc
     5641 gctccatcag catattcatg tagaaattga tgatctcatc attgagccaa ttcagatggt
     5701 ttagagtttg aatatctttg cgtgtaatgg tcaggcgaaa tgcttcactg agaacttcat
     5761 cctgattccc attacgaaat acattcttta tttctttctc catttcctct gtaatttcag
     5821 gaaattccat atggctgccg cgcggcacca ggccgctgct gtgatgatga tgatgatggc
     5881 tgctgcccat ggtatatctc cttcttaaag ttaaacaaaa ttatttctag aggggaattg
     5941 ttatccgctc acaattcccc tatagtgagt cgt
//
