LOCUS       sequence-379509-        5872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express C57D/C146D mutant SOD1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5872)
  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 5872)
  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: pcDNA3.1-SOD1 C57D/C146D
FEATURES             Location/Qualifiers
     source          1..5872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(180..199)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        371..750
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        751..954
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     905..925
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     999..1018
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        999..1017
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1599..1616)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1605..1829
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1875..2303
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1962..1981)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2172..2193
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2312..2332)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2317..2646
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2497..2632
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2559..2578
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2713..3507
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWVQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     3377..3396
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3681..3802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3718..3737)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3772..3791
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3851..3867)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3851..3867)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3864..3886)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3875..3891
                     /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)."
     promoter        complement(join(3899..3905,3906..3923,3924..3929))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3944..3965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4082..4099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4253..4841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4333..4352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5012..5803,5804..5872))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5635..5654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
      181 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
      241 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
      301 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
      361 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      421 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      481 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      541 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      601 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      661 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      721 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      781 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      841 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      901 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
      961 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     1021 acccaagctg gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag
     1081 tgtggtggaa ttccatggcg acgaaggccg tgtgcgtgct gaagggcgac ggcccagtgc
     1141 agggcatcat caatttcgag cagaaggaaa gtaatggacc agtgaaggtg tggggaagca
     1201 ttaaaggact gactgaaggc ctgcatggat tccatgttca tgagtttgga gataatacag
     1261 caggcgatac cagtgcaggt cctcacttta atcctctatc cagaaaacac ggtgggccaa
     1321 aggatgaaga gaggcatgtt ggagacttgg gcaatgtgac tgctgacaaa gatggtgtgg
     1381 ccgatgtgtc tattgaagat tctgtgatct cactctcagg agaccattgc atcattggcc
     1441 gcacactggt ggtccatgaa aaagcagatg acttgggcaa aggtggaaat gaagaaagta
     1501 caaagacagg aaacgctgga agtcgtttgg ctgatggtgt aattgggatc gcccaataac
     1561 gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     1621 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     1681 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     1741 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     1801 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     1861 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     1921 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     1981 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2041 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2101 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2161 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2221 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2281 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     2341 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2401 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2461 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2521 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2581 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2641 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     2701 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     2761 gagaggctat tcggctatga ctgggtacaa cagacaatcg gctgctctga tgccgccgtg
     2821 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     2881 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     2941 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3001 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3061 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3121 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3181 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3241 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3301 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3361 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3421 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3481 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     3541 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     3601 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     3661 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     3721 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3781 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     3841 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3901 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3961 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4021 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4081 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4141 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4201 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4261 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4321 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4381 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4441 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4501 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4561 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4621 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4681 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4741 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4801 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4861 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4921 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4981 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5041 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5101 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5161 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5221 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5281 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5341 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5401 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5461 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5521 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5581 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5641 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5701 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5761 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5821 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc at
//
