LOCUS       sequence-138323-        6013 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 6013)
  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 6013)
  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-SENP2 (catalytic domain)
FEATURES             Location/Qualifiers
     source          1..6013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(85..104)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        310..387
                     /gene="lacI"
                     /label=lacI promoter
     CDS             388..1470
                     /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(407..426)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2279..2470
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2464..2486)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2572..2711
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2627..2646
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2726..2743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2897..3485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2977..2996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3596..3615)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3607..4422
                     /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             3607..4422
                     /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(3680..3699)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(4514..4969)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4651..4672)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4863..4882
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(5006..5053)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     5049..5067
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(5120..5137)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5871..5888)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(5898..5915)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             5934..5956
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5971..5995
                     /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(5995..6013,1))
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(join(5996..6013,1))
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aatttcgcgg gatcgagatc tcgatcctct acgccggacg catcgtggcc ggcatcaccg
       61 gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg
      121 ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg
      181 ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca
      241 acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc
      301 gagatcccgg acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
      361 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
      421 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
      481 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
      541 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
      601 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
      661 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
      721 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
      781 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
      841 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
      901 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
      961 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     1021 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     1081 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     1141 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     1201 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     1261 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     1321 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     1381 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     1441 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac
     1501 tcattaggca ccgggatctc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     1561 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     1621 actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc gctttcgctg
     1681 gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca
     1741 agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg
     1801 catggcggcc ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg
     1861 gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga
     1921 ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt
     1981 ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc ggatctgcat
     2041 cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga agcgctggca
     2101 ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt gtttaccctc
     2161 acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga gcatcctctc
     2221 tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg aggcatcagt
     2281 gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac
     2341 gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct
     2401 tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga
     2461 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2521 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     2581 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     2641 attgtactga gagtgcacca ttgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2701 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2761 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2821 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2881 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2941 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3001 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3061 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3121 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3181 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3241 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3301 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     3361 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3421 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3481 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3541 gttaagggat tttggtcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     3601 ggtgttatga gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac
     3661 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     3721 acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     3781 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     3841 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     3901 actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     3961 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     4021 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     4081 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     4141 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     4201 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     4261 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     4321 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     4381 aataaattgc agtttcattt gatgctcgat gagtttttct aagaattaat tcatgagcgg
     4441 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4501 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4561 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4621 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4681 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4741 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4801 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4861 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4921 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     4981 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
     5041 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
     5101 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
     5161 cgacggagct cgaattcgga tcctcacagc aactgctgat gaaggatttc ccacaccatc
     5221 ttcttccgga agagaggcat ctggtgctga gtaaatgtga taggtttgtc cctagaaata
     5281 taatctgcat atttacaagt aaacattcca caatcactcc cattcagctg ttgaggaatc
     5341 tcgtgtggtt tcatgctgtg atgggtccac tctaaaagat tcagatcact atttcttttg
     5401 gtcttacttt catcctgtaa atactgaagg agaatctcac agatcctgtg gcccttttgt
     5461 cccatagaat ccagatattt aagacacttt tttcttaggt caatcaccac caggctccaa
     5521 tgtaccttcc gatgaatagg caccagaata atttcttgtt caaagagatt tacccctttg
     5581 gtccatcgtt tcactgcttg gtaaccccca gactttaatt taggatagaa gaaagtactg
     5641 aatacatgaa gtgctggata gccttgcttt ttatttcttt ccaccagaag attcatgtaa
     5701 aaattaatga cttcatcatt gagccagtga tagttcttta atgtctgaat atctcctcga
     5761 gtaattcgca atttgaaagc actacttagg atttcatcct gtgggccatg gcctagggca
     5821 ttactgattt ccttttccat gtcctctgta agttcaagga gatccatatg gctgccgcgc
     5881 ggcaccaggc cgctgctgtg atgatgatga tgatggctgc tgcccatggt atatctcctt
     5941 cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca attcccctat
     6001 agtgagtcgt att
//
