LOCUS       sequence-332852-        7090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7090)
  AUTHORS   Dirac-Svejstrup AB, Walker J, Faull P, Encheva V, Akimov V, Puglia 
            M, Perkins D, Kumper S, Hunjan SS, Blagoev B, Snijders AP, Powell 
            DJ, Svejstrup JQ
  TITLE     DDI2 Is a Ubiquitin-Directed Endoprotease Responsible for Cleavage 
            of Transcription Factor NRF1.
  JOURNAL   Mol Cell. 2020 Jul 16;79(2):332-341.e7. doi: 
            10.1016/j.molcel.2020.05.035. Epub 2020 Jun 9.
  PUBMED    32521225
REFERENCE   2  (bases 1 to 7090)
  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: 
            "10.1016/j.molcel.2020.05.035"; journalName: "Mol Cell"; date: 
            "2020-07-16- 16"; volume: "79"; issue: "2"; pages: "332-341.e7"
COMMENT     Sequence Label: His-SUMO-Rad23A_pET11a
FEATURES             Location/Qualifiers
     source          1..7090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             101..118
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             128..145
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1591..1609)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1605..1652
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1827..1832,1833..1849,1850..1855))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1882..1900)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1968..2072
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2073..2141,2142..2933)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2291..2310)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3104..3692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3593..3612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3846..3863
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3878..4017
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3943..3962)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4103..4125
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4119..4310)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5622..6704)
                     /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     6666..6685
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6705..6782)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6988..7007
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gggcagcagc catcatcatc atcatcatgg
      121 tagcggtctg gttccgcgtg gtagcgcgag catgagcgac agcgaagtga atcaagaagc
      181 gaagccggaa gtgaagccgg aagtgaagcc ggaaacccac atcaacctga aggttagcga
      241 tggcagcagc gaaatcttct ttaagattaa gaaaaccacc ccgctgcgtc gtctgatgga
      301 agcgttcgcg aagcgtcagg gcaaagagat ggacagcctg cgttttctgt acgatggtat
      361 ccgtattcag gcggaccaaa ccccggaaga cctggatatg gaggacaacg atatcattga
      421 ggcgcaccgt gaacagatcg gtggtggcat ggcggtgacc attaccctga agaccctgca
      481 gcaacagacc ttcaaaatcc gtatggagcc ggatgaaacc gttaaggtgc tgaaggaaaa
      541 aattgaggcg gaaaaaggtc gtgacgcgtt tccggttgcg ggccagaagc tgatctacgc
      601 gggtaaaatt ctgagcgacg atgtgccgat ccgtgactat cgtattgatg agaagaactt
      661 cgtggttgtg atggttacca agaccaaggc gggtcaaggt accagcgctc cgccggaagc
      721 gagcccgacc gcggcgccgg agagcagcac cagctttccg ccggcgccga ccagcggtat
      781 gagccacccg ccgccggcgg cgcgtgaaga taaaagcccg agcgaagaga gcgcgccgac
      841 caccagcccg gagagcgtta gcggtagcgt gccgagcagc ggtagcagcg gtcgtgagga
      901 agacgcggcg agcaccctgg ttaccggcag cgagtacgaa accatgctga ccgaaatcat
      961 gagcatgggt tatgagcgtg aacgtgttgt ggcggcgctg cgtgcgagct acaacaaccc
     1021 gcaccgtgcg gtggagtatc tgctgaccgg tattccgggc agcccggagc cggaacatgg
     1081 cagcgttcag gaaagccaag tgagcgagca gccggcgacc gaggcggcgg gtgaaaaccc
     1141 gctggagttc ctgcgtgacc aaccgcagtt tcaaaacatg cgtcaagtga tccaacaaaa
     1201 cccggcgctg ctgccggcgc tgctgcaaca gctgggccag gaaaacccgc aactgctgca
     1261 acagatcagc cgtcaccagg agcaattcat tcagatgctg aacgaaccgc cgggcgagct
     1321 ggcggacatc agcgatgttg agggtgaagt gggcgcgatt ggcgaggaag cgccgcaaat
     1381 gaactacatc caggttaccc cgcaagagaa ggaagcgatt gaacgtctga aagcgctggg
     1441 tttcccggag agcctggtga ttcaggcgta ttttgcgtgc gagaaaaatg agaacctggc
     1501 ggcgaacttc ctgctgagcc agaactttga cgacgaatag ggatccggct gctaacaaag
     1561 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     1621 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc
     1681 ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac
     1741 ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta
     1801 gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg
     1861 agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     1921 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1981 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2041 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2101 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2161 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2221 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2281 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     2341 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2401 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2461 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2521 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2581 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     2641 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2701 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2761 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2821 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2881 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2941 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3001 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3061 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3121 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3181 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3241 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3301 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3361 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3421 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3481 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3541 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3601 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3661 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3721 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3781 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3841 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     3901 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
     3961 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     4021 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4081 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4141 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     4201 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     4261 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     4321 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     4381 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     4441 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     4501 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     4561 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     4621 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     4681 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     4741 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
     4801 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg
     4861 gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat
     4921 ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc
     4981 gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt
     5041 tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca atgatcgaag
     5101 ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct
     5161 gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca
     5221 taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt
     5281 cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     5341 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     5401 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     5461 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     5521 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     5581 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     5641 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5701 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     5761 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     5821 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     5881 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     5941 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     6001 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     6061 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     6121 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     6181 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     6241 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     6301 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     6361 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     6421 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     6481 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     6541 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     6601 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     6661 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     6721 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     6781 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     6841 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     6901 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     6961 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     7021 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat
     7081 cccgcgaaat
//
