LOCUS       sequence-270222-        5828 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 5828)
  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 5828)
  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..5828
                     /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        1228..1332
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1333..1401,1402..2193)
                     /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(1551..1570)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2364..2952
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2853..2872
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3106..3123
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3138..3277
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3203..3222)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3363..3385
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3379..3570)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4379..5461)
                     /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     5423..5442
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5462..5539)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5745..5764
                     /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 gtttacaatt
     1201 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1261 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1321 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     1381 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     1441 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     1501 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     1561 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     1621 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     1681 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     1741 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     1801 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     1861 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     1921 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     1981 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2041 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2101 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2161 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     2221 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2281 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2341 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2401 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2461 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     2521 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2581 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2641 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2701 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2761 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2821 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2881 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2941 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3001 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3061 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3121 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3181 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     3241 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     3301 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3361 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     3421 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     3481 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     3541 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     3601 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     3661 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     3721 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     3781 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     3841 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     3901 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     3961 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     4021 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     4081 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     4141 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     4201 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     4261 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     4321 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     4381 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4441 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     4501 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     4561 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     4621 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     4681 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     4741 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     4801 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     4861 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     4921 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     4981 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     5041 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     5101 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     5161 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     5221 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     5281 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     5341 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     5401 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     5461 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     5521 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     5581 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     5641 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     5701 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     5761 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     5821 aggatcga
//
