LOCUS       sequence-274741-        5827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmids containing human A4V/C6S SOD1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5827)
  AUTHORS   Chantadul V, Wright GSA, Amporndanai K, Shahid M, Antonyuk SV, 
            Washbourn G, Rogers M, Roberts N, Pye M, O'Neill PM, Hasnain SS
  TITLE     Ebselen as template for stabilization of A4V mutant dimer for motor 
            neuron disease therapy.
  JOURNAL   Commun Biol. 2020 Mar 5;3(1):97. doi: 10.1038/s42003-020-0826-3.
  PUBMED    32139772
REFERENCE   2  (bases 1 to 5827)
  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: "2020-03-5"; volume: "3(1):97. doi"; pages: " 
            10.1038/s42003-020-0826-3"
COMMENT     Sequence Label: pET303C-hSOD1 A4V/C6S
FEATURES             Location/Qualifiers
     source          1..5827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    39..63
                     /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             79..99
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             578..595
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(648..666)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      662..709
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      746..1201
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(833..852)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1043..1064
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1227..1331
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1332..1400,1401..2192)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1550..1569)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2363..2951
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2852..2871
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3105..3122
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3137..3276
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3202..3221)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3362..3384
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3378..3569)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4378..5460)
                     /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     5422..5441
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5461..5538)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5744..5763
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
       61 tcccctctag taataatttt gtttaacttt aagaaggagg tctagaatgg cgacgaaggt
      121 cgtgagcgtg ctgaagggcg acggcccagt gcagggcatc atcaatttcg agcagaagga
      181 aagtaatgga ccagtgaagg tgtggggaag cattaaagga ctgactgaag gcctgcatgg
      241 attccatgtt catgagtttg gagataatac agcaggctgt accagtgcag gtcctcactt
      301 taatcctcta tccagaaaac acggtgggcc aaaggatgaa gagaggcatg ttggagactt
      361 gggcaatgtg actgctgaca aagatggtgt ggccgatgtg tctattgaag attctgtgat
      421 ctcactctca ggagaccatt gcatcattgg ccgcacactg gtggtccatg aaaaagcaga
      481 tgacttgggc aaaggtggaa atgaagaaag tacaaagaca ggaaacgctg gaagtcgttt
      541 ggcttgtggt gtaattggga tcgcccaata actcgagcac caccaccacc accactgaga
      601 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
      661 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
      721 aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
      781 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      841 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      901 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
      961 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1021 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1081 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1141 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1201 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1261 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1321 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1381 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1441 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1501 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1561 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1621 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1681 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1741 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1801 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1861 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1921 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1981 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2041 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2101 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2161 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2221 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2281 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2341 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2401 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2461 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     2521 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2581 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2641 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2701 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2761 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2821 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2881 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2941 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3001 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3061 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3121 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3181 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     3241 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     3301 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3361 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     3421 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     3481 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     3541 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     3601 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     3661 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     3721 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     3781 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     3841 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     3901 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     3961 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     4021 atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     4081 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     4141 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     4201 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     4261 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     4321 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     4381 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4441 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     4501 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     4561 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     4621 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     4681 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     4741 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     4801 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     4861 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     4921 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     4981 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     5041 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     5101 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     5161 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     5221 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     5281 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     5341 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     5401 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     5461 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     5521 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     5581 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     5641 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     5701 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     5761 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     5821 ggatcga
//
