LOCUS       sequence-378588-        5855 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression plasmid for human WT SOD1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5855)
  AUTHORS   Switzer CH, Kasamatsu S, Ihara H, Eaton P
  TITLE     SOD1 is an essential H(2)S detoxifying enzyme.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Jan 17;120(3):e2205044120. doi: 
            10.1073/pnas.2205044120. Epub 2023 Jan 11.
  PUBMED    36630448
REFERENCE   2  (bases 1 to 5855)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2023-01-17"; volume: "120(3):e2205044120. 
            doi"; pages: " 10.1073/pnas.2205044120. Epub 2023 Jan 11"
COMMENT     Sequence Label: hSOD1-FLAG
FEATURES             Location/Qualifiers
     source          1..5855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(66..85)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        257..636
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        637..840
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     791..811
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     885..904
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        885..903
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      957..966
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1425..1448
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1468..1485)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1474..1698
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1744..2172
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1831..1850)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2041..2062
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2181..2201)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2186..2515
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2366..2501
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2428..2447
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2582..3376
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2636..2655)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3246..3265
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3550..3671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3587..3606)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3641..3660
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3720..3736)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3720..3736)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3733..3755)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3744..3760
                     /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(3768..3774,3775..3792,3793..3798))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3813..3834
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3951..3968)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4122..4710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4202..4221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4881..5672,5673..5741))
                     /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     5504..5523
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5742..5846)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
       61 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
      121 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
      181 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
      241 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      301 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      361 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      421 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      481 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      541 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      601 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      661 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      721 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      781 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      841 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      901 agggagaccc aagctggcta gcgtttaaac ttaagcttgg taccgagctc ggatccgcca
      961 ccatggcgac gaaggccgtg tgcgtgctga agggcgacgg cccagtgcag ggcatcatca
     1021 atttcgagca gaaggaaagt aatggaccag tgaaggtgtg gggaagcatt aaaggactga
     1081 ctgaaggcct gcatggattc catgttcatg agtttggaga taatacagca ggctgtacca
     1141 gtgcaggtcc tcactttaat cctctatcca gaaaacacgg tgggccaaag gatgaagaga
     1201 ggcatgttgg agacttgggc aatgtgactg ctgacaaaga tggtgtggcc gatgtgtcta
     1261 ttgaagattc tgtgatctca ctctcaggag accattgcat cattggccgc acactggtgg
     1321 tccatgaaaa agcagatgac ttgggcaaag gtggaaatga agaaagtaca aagacaggaa
     1381 acgctggaag tcgtttggct tgtggtgtaa ttgggatcgc ccaagattac aaggatgacg
     1441 acgataagtg ataaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     1501 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     1561 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     1621 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     1681 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     1741 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1801 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1861 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1921 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1981 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2041 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2101 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2161 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2221 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2281 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2341 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2401 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     2461 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     2521 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     2581 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2641 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     2701 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     2761 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     2821 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     2881 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     2941 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3001 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3061 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     3121 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3181 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3241 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3301 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3361 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3421 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3481 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3541 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3601 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3661 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     3721 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3781 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3841 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3901 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3961 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4021 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4081 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4141 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4201 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4261 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4321 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4381 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4441 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4501 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4561 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4621 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4681 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4741 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4801 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4861 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4921 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4981 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5041 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5101 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5161 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5221 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5281 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5341 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5401 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5461 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5521 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5581 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5641 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5701 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5761 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5821 aaaaataaac aaataggggt tccgcgcaca tttcc
//
