LOCUS       sequence-379505-        5138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express C57A/C146A 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 C57A/C146A
FEATURES             Location/Qualifiers
     source          1..5138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(248..676)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(358..379)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     570..589
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(714..733)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     874..896
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        974..1051
                     /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             1052..2134
                     /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(1071..1090)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    2403..2419
                     /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     2408..2430
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             2452..2469
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2491..2511
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3033..3050)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3203..3289
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3381..3408
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3427..3518
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3519..3587,3588..4379)
                     /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(3737..3756)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4550..5138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5039..5058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataatt
      241 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
      301 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
      361 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
      421 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
      481 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
      541 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
      601 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
      661 ccgctacagg gcgcgtccca ttcgccattc aggctgctat ggtgcactct cagtacaatc
      721 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
      781 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
      841 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
      901 caccgtcatc accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc
      961 atgcatttac gttgacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg
     1021 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     1081 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     1141 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     1201 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     1261 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     1321 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     1381 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     1441 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     1501 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     1561 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     1621 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     1681 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     1741 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     1801 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     1861 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     1921 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     1981 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     2041 ggcacccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     2101 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     2161 gcgaattgat ctggtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     2221 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     2281 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     2341 ctggcaaata ttctgaaatg agctgttgac aattaatcat ccggtccgta taatctgtgg
     2401 aattgtgagc ggataacaat ttcacacagg aaacagacca tgtcgtacta ccatcaccat
     2461 caccatcacg attacgatat cccaacgacc gaaaacctgt attttcaggg cgccatggat
     2521 ccggaattca tggcgacgaa ggccgtgtgc gtgctgaagg gcgacggccc agtgcagggc
     2581 atcatcaatt tcgagcagaa ggaaagtaat ggaccagtga aggtgtgggg aagcattaaa
     2641 ggactgactg aaggcctgca tggattccat gttcatgagt ttggagataa tacagcaggc
     2701 gctaccagtg caggtcctca ctttaatcct ctatccagaa aacacggtgg gccaaaggat
     2761 gaagagaggc atgttggaga cttgggcaat gtgactgctg acaaagatgg tgtggccgat
     2821 gtgtctattg aagattctgt gatctcactc tcaggagacc attgcatcat tggccgcaca
     2881 ctggtggtcc atgaaaaagc agatgacttg ggcaaaggtg gaaatgaaga aagtacaaag
     2941 acaggaaacg ctggaagtcg tttggctgct ggtgtaattg ggatcgccca ataaaagctt
     3001 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga
     3061 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc
     3121 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg
     3181 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     3241 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     3301 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     3361 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
     3421 aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3481 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3541 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3601 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3661 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3721 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     3781 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3841 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3901 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3961 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4021 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4081 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4141 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4201 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4261 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4321 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4381 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4441 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4501 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4561 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4621 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4681 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4741 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4801 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4861 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4921 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4981 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5041 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5101 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaa
//
