LOCUS       sequence-379503-        5138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express C57D/C146D mutant SOD1 in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5138)
  AUTHORS   Baek Y, Woo TG, Ahn J, Lee D, Kwon Y, Park BJ, Ha NC
  TITLE     Structural analysis of the overoxidized Cu/Zn-superoxide dismutase 
            in ROS-induced ALS filament formation.
  JOURNAL   Commun Biol. 2022 Oct 12;5(1):1085. doi: 10.1038/s42003-022-04017-0.
  PUBMED    36224351
REFERENCE   2  (bases 1 to 5138)
  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: "Commun 
            Biol."; date: "2022-10-12"; volume: "5(1):1085. doi"; pages: " 
            10.1038/s42003-022-04017-0"
COMMENT     Sequence Label: pProEx-HTa-SOD1 C57D/C146D
FEATURES             Location/Qualifiers
     source          1..5138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..57)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     198..220
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        298..375
                     /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."
     CDS             376..1458
                     /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(395..414)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    1727..1743
                     /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     1732..1754
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1776..1793
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1815..1835
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2357..2374)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2527..2613
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2705..2732
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2751..2842
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2843..2911,2912..3703)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3061..3080)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3874..4462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4363..4382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4616..4633
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4710..5138)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4820..4841)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5032..5051
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cccattcgcc attcaggctg ctatggtgca ctctcagtac aatctgctct gatgccgcat
       61 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
      121 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
      181 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
      241 acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag cggcatgcat ttacgttgac
      301 accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
      361 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
      421 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
      481 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
      541 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
      601 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
      661 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
      721 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
      781 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
      841 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
      901 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
      961 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
     1021 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
     1081 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
     1141 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat
     1201 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg
     1261 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
     1321 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcacc caatacgcaa
     1381 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     1441 ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagcgcgaat tgatctggtt
     1501 tgacagctta tcatcgactg cacggtgcac caatgcttct ggcgtcaggc agccatcgga
     1561 agctgtggta tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca
     1621 ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga
     1681 aatgagctgt tgacaattaa tcatccggtc cgtataatct gtggaattgt gagcggataa
     1741 caatttcaca caggaaacag accatgtcgt actaccatca ccatcaccat cacgattacg
     1801 atatcccaac gaccgaaaac ctgtattttc agggcgccat ggatccggaa ttcatggcga
     1861 cgaaggccgt gtgcgtgctg aagggcgacg gcccagtgca gggcatcatc aatttcgagc
     1921 agaaggaaag taatggacca gtgaaggtgt ggggaagcat taaaggactg actgaaggcc
     1981 tgcatggatt ccatgttcat gagtttggag ataatacagc aggcgatacc agtgcaggtc
     2041 ctcactttaa tcctctatcc agaaaacacg gtgggccaaa ggatgaagag aggcatgttg
     2101 gagacttggg caatgtgact gctgacaaag atggtgtggc cgatgtgtct attgaagatt
     2161 ctgtgatctc actctcagga gaccattgca tcattggccg cacactggtg gtccatgaaa
     2221 aagcagatga cttgggcaaa ggtggaaatg aagaaagtac aaagacagga aacgctggaa
     2281 gtcgtttggc tgatggtgta attgggatcg cccaataaaa gcttggctgt tttggcggat
     2341 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     2401 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     2461 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     2521 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     2581 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     2641 aagcaacggc ccggagggtg gcgggcagga cgtccgccat aaactgccag gcatcaaatt
     2701 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     2761 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2821 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2881 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2941 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3001 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3061 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     3121 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3181 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3241 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3301 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3361 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3421 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3481 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3541 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3601 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3661 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3721 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3781 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3841 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3901 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3961 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4021 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4081 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4141 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4201 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4261 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4321 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4381 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4441 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4501 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4561 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4621 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     4681 cggtatttca caccgcataa ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4741 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4801 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     4861 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     4921 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     4981 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5041 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5101 cacacccgcc gcgcttaatg cgccgctaca gggcgcgt
//
