LOCUS       sequence-379507-        5138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express wild-type 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 WT
FEATURES             Location/Qualifiers
     source          1..5138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(72..91)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     232..254
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        332..409
                     /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             410..1492
                     /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(429..448)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    1761..1777
                     /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     1766..1788
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1810..1827
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1849..1869
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2391..2408)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2561..2647
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2739..2766
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2785..2876
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2877..2945,2946..3737)
                     /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(3095..3114)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3908..4496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4397..4416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4650..4667
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4744..5138,1..34))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4854..4875)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5066..5085
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgctatgg
       61 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
      121 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
      181 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
      241 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc
      301 gcgcgaaggc gaagcggcat gcatttacgt tgacaccatc gaatggtgca aaacctttcg
      361 cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt
      421 aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt
      481 gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga
      541 gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat
      601 tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa
      661 atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt
      721 cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat
      781 taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc
      841 ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga
      901 agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct
      961 gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata
     1021 tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc
     1081 cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt
     1141 tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt
     1201 tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc
     1261 gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt
     1321 gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt
     1381 gaaaagaaaa accaccctgg cacccaatac gcaaaccgcc tctccccgcg cgttggccga
     1441 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     1501 caattaatgt aagttagcgc gaattgatct ggtttgacag cttatcatcg actgcacggt
     1561 gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt
     1621 aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc
     1681 gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcc
     1741 ggtccgtata atctgtggaa ttgtgagcgg ataacaattt cacacaggaa acagaccatg
     1801 tcgtactacc atcaccatca ccatcacgat tacgatatcc caacgaccga aaacctgtat
     1861 tttcagggcg ccatggatcc ggaattcatg gcgacgaagg ccgtgtgcgt gctgaagggc
     1921 gacggcccag tgcagggcat catcaatttc gagcagaagg aaagtaatgg accagtgaag
     1981 gtgtggggaa gcattaaagg actgactgaa ggcctgcatg gattccatgt tcatgagttt
     2041 ggagataata cagcaggctg taccagtgca ggtcctcact ttaatcctct atccagaaaa
     2101 cacggtgggc caaaggatga agagaggcat gttggagact tgggcaatgt gactgctgac
     2161 aaagatggtg tggccgatgt gtctattgaa gattctgtga tctcactctc aggagaccat
     2221 tgcatcattg gccgcacact ggtggtccat gaaaaagcag atgacttggg caaaggtgga
     2281 aatgaagaaa gtacaaagac aggaaacgct ggaagtcgtt tggcttgtgg tgtaattggg
     2341 atcgcccaat aaaagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag
     2401 attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg
     2461 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt
     2521 gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
     2581 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     2641 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     2701 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
     2761 cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc
     2821 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2881 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2941 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3001 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3061 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     3121 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3181 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3241 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3301 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3361 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3421 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3481 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3541 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3601 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3661 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3721 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3781 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3841 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3901 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3961 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4021 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4081 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4141 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4201 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4261 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4321 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4381 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4441 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4501 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4561 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4621 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4681 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataattttgt
     4741 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4801 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4861 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4921 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4981 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     5041 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     5101 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccaca
//
