LOCUS       sequence-398273-        4068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of HsPI31-1-151.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4068)
  AUTHORS   Kraus F, Goodall EA, Smith IR, Jiang Y, Paoli JC, Adolf F, Zhang J, 
            Paulo JA, Schulman BA, Harper JW
  TITLE     PARK15/FBXO7 is dispensable for PINK1/Parkin mitophagy in iNeurons 
            and HeLa cell systems.
  JOURNAL   EMBO Rep. 2023 Aug 3;24(8):e56399. doi: 10.15252/embr.202256399. 
            Epub 2023 Jun 19.
  PUBMED    37334901
REFERENCE   2  (bases 1 to 4068)
  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: "EMBO Rep.";
            date: "2023-08-3"; volume: "24(8):e56399. doi"; pages: " 
            10.15252/embr.202256399. Epub 2023 Jun 19"
COMMENT     Sequence Label: pRSF1b-His8-TEV-PI31-1-151
FEATURES             Location/Qualifiers
     source          1..4068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             83..106
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             107..127
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             605..649
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(687..705)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      701..748
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(978..1793)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(978..1793)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1701..1720
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1794..1885)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1901..2650)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(1911..1928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2151..2170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2846..3928)
                     /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     3890..3909
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3929..4006)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(4052..4068,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4052..4068,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcaccatcac catcacgagg atctgtactt
      121 tcagtctatg gcgggcctgg aggtactgtt cgcatcggca gcgccggcca tcacctgcag
      181 gcaggacgcg ctcgtctgct tcttgcattg ggaagtggtg acacacggtt actgcggctt
      241 gggtgtcggt gaccagccgg gtcccaatga taagaagtca gaactgctgc cagctgggtg
      301 gaacaacaat aaagacctgt atgtcctccg gtatgagtat aaggatgggt ccagaaagct
      361 ccttgtgaaa gccatcaccg tggagagcag catgatcctc aatgtgctgg aatatggctc
      421 acagcaagtg gcagacttga ccctgaactt ggatgattat atcgatgcag aacacctggg
      481 tgacttccac aggacctaca agaacagtga ggagcttcgg tctcgtattg tgtctggaat
      541 catcacacct atccatgagc agtgggaaaa ggctaatgta tgataaggta ccctcgagtc
      601 tggtaaagaa accgctgctg cgaaatttga acgccagcac atggactcgt ctactagcgc
      661 agcttaatta acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
      721 ctctaaacgg gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt
      781 cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta
      841 acgaccctgc cctgaaccga cgacaagctg acgaccgggt ctccgcaagt ggcacttttc
      901 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      961 cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc
     1021 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     1081 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     1141 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     1201 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     1261 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     1321 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc ggtcgctgtt aaaaggacaa
     1381 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     1441 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     1501 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     1561 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     1621 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     1681 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     1741 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct catactcttc
     1801 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1861 gaatgtattt agaaaaataa acaaataggc atgcagcgct cttccgcttc ctcgctcact
     1921 gactcgctac gctcggtcgt tcgactgcgg cgagcggtgt cagctcactc aaaagcggta
     1981 atacggttat ccacagaatc aggggataaa gccggaaaga acatgtgagc aaaaagcaaa
     2041 gcaccggaag aagccaacgc cgcaggcgtt tttccatagg ctccgccccc ctgacgagca
     2101 tcacaaaaat cgacgctcaa gccagaggtg gcgaaacccg acaggactat aaagatacca
     2161 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2221 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgttg
     2281 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2341 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2401 cgacttatcg ccactggcag cagccattgg taactgattt agaggacttt gtcttgaagt
     2461 tatgcacctg ttaaggctaa actgaaagaa cagattttgg tgagtgcggt cctccaaccc
     2521 acttaccttg gttcaaagag ttggtagctc agcgaacctt gagaaaacca ccgttggtag
     2581 cggtggtttt tctttattta tgagatgatg aatcaatcgg tctatcaagt caacgaacag
     2641 ctattccgtt actctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca
     2701 gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc accggaagga
     2761 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     2821 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2881 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     2941 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     3001 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     3061 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     3121 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     3181 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     3241 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     3301 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     3361 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     3421 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     3481 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     3541 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     3601 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     3661 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     3721 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     3781 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     3841 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     3901 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     3961 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     4021 tctcccttat gcgactcctg cattaggaaa ttaatacgac tcactata
//
