LOCUS       sequence-332235-        7754 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 7754)
  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 7754)
  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: Nrf1-Myc pcDNA3.1(+)-C-Myc (NAWLVH106-AAAAAA)
FEATURES             Location/Qualifiers
     source          1..7754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3062..3091
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3111..3128)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3117..3341
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3387..3815
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3474..3493)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3684..3705
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3824..3844)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3829..4158
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4009..4144
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4225..5019
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4279..4298)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4889..4908
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5193..5314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5230..5249)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5284..5303
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5363..5379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5363..5379)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5376..5398)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5387..5403
                     /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)."
     promoter        complement(join(5411..5417,5418..5435,5436..5441))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5456..5477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5594..5611)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5765..6353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5845..5864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6524..7315,7316..7384))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7147..7166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7385..7489)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7564..7583)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc gaattctgca gatatccagc
      721 acagtggcgg ccgctcgaga tgctttctct gaagaaatac ttaacggaag gacttctcca
      781 gttcaccatt ctgctgagtt tgattggggt acgggtggac gtggatactt acctgacctc
      841 acagcttccc ccactccggg agatcatcct ggggcccagt tctgcctata ctcagaccca
      901 gttccacaac ctgaggaata ccttggatgg ctatggtatc caccccaaga gcatagacct
      961 ggacaattac ttcactgccc ggcggctcct cagtcaggtg agggccctgg acaggttcca
     1021 ggtgccaacc actgaggtag ctgccgcggc ggctgcccga gacccagagg ggtctgtctc
     1081 tggcagtcag cccaactcag gcctcgccct cgagagttcc agtggcctcc aagatgtgac
     1141 aggcccagac aacggggtgc gagaaagcga aacggagcag ggattcggtg aagatttgga
     1201 ggatttgggg gctgtagccc ccccagtcag tggagactta accaaagagg acatagatct
     1261 gattgacatc ctttggcgac aggatattga tctgggggct gggcgtgagg tttttgacta
     1321 tagtcaccgc cagaaggagc aggatgtgga gaaggagctg cgagatggag gcgagcagga
     1381 cacctgggca ggcgagggcg cggaagctct ggcacggaac ctgctagtgg atggagagac
     1441 tggggagagc ttccctgcac aggtgcctag tggggaggac cagacggccc tgtccctgga
     1501 agagtgcctt aggctgctgg aagccacctg cccctttggg gagaatgctg agtttccagc
     1561 agacatttcc agcataacag aagcagtgcc tagtgagagt gagccccctg ctcttcaaaa
     1621 caacctcttg tctcctcttc tgaccgggac agagtcacca tttgatttgg aacagcagtg
     1681 gcaagatctc atgtccatca tggaaatgca ggccatggaa gtgaacacat cagcaagtga
     1741 aatcctgtac agtgcccctc ctggagaccc actgagcacc aactacagcc ttgcccccaa
     1801 cactcccatc aatcagaatg tcagcctgca tcaggcgtcc ctggggggct gcagccagga
     1861 cttcttactc ttcagccccg aggtggaaag cctgcctgtg gccagtagct ccacgctgct
     1921 cccgttggcc cccagcaatt ctaccagcct caactccacc ttcggctcca ccaacctgac
     1981 agggctcttc tttccacccc agctcaatgg cacagccaat gacacagcag gcccagagct
     2041 gcctgaccct ttggggggtc tgttagatga agctatgttg gatgagatca gccttatgga
     2101 cctggccatt gaagaaggct ttaaccctgt gcaggcctcc cagctggagg aggaatttga
     2161 ctctgactca ggcctttcct tagactcgag ccatagccct tcttccctaa gcagctctga
     2221 aggcagttct tcctcttctt cctcctcctc ttcctcttct tcctctgctt cttcctctgc
     2281 ctcttcctcc ttttctgagg aaggtgcggt tggctacagc tctgactctg agaccctgga
     2341 tctggaagag gccgagggtg ctgtgggcta ccagcctgag tattccaagt tctgccgcat
     2401 gagctaccag gatccagctc agctctcatg cctgccctac ctggagcacg tgggccacaa
     2461 ccacacatac aacatggcac ccagtgccct ggactcagcc gacctgccac cacccagtgc
     2521 cctcaagaaa ggcagcaagg agaagcaggc tgacttcctg gacaagcaga tgagccggga
     2581 tgagcaccga gcccgagcca tgaagatccc tttcaccaat gacaaaatca tcaacctgcc
     2641 tgtggaggag ttcaatgaac tgctgtccaa ataccagttg agtgaagccc agctgagcct
     2701 catccgagac atccggcgcc ggggcaagaa caagatggcg gcgcagaact gccgcaagcg
     2761 caagctggac accatcctga atctggagcg tgatgtggag gacctgcagc gtgacaaagc
     2821 ccggctgctg cgggagaaag tggagttcct gcgctccctg cgacagatga agcagaaggt
     2881 ccagagcctg taccaggagg tgtttgggcg gctgcgagat gagaacggac gaccctactc
     2941 gcccagtcag tatgcgctcc agtacgccgg ggacggcagt gtcctcctca tcccccgcac
     3001 gatggccgac cagcaggccc ggcggcagga gaggaagcca aaggaccgga gaaaggggcc
     3061 cgagcagaaa ctcatctctg aagaggatct gtgataaacc cgctgatcag cctcgactgt
     3121 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3181 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3241 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3301 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3361 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3421 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3481 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3541 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3601 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3661 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3721 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3781 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3841 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3901 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3961 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4021 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4081 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4141 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4201 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4261 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4321 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4381 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4441 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4501 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4561 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4621 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4681 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4741 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4801 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4861 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4921 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4981 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5041 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5101 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5161 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5221 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5281 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5341 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5401 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5461 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5701 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5761 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5821 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5881 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5941 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6001 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6061 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6121 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6181 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6241 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6301 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6361 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6421 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6481 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6541 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6601 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6661 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6721 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6781 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6841 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6901 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6961 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7021 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7081 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7141 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7201 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7261 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7321 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7381 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7441 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7501 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     7561 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7621 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     7681 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     7741 cagatatacg cgtt
//
