LOCUS       sequence-197770-        6229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Rat Parkin with residues 86-130 deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6229)
  AUTHORS   Sauve V, Lilov A, Seirafi M, Vranas M, Rasool S, Kozlov G, Sprules 
            T, Wang J, Trempe JF, Gehring K
  TITLE     A Ubl/ubiquitin switch in the activation of Parkin.
  JOURNAL   EMBO J. 2015 Oct 14;34(20):2492-505. doi: 10.15252/embj.201592237. 
            Epub 2015 Aug 7.
  PUBMED    26254305
REFERENCE   2  (bases 1 to 6229)
  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"; doi: 
            "10.15252/embj.201592237"; journalName: "EMBO J"; date: "2015-10-14-
            14"; volume: "34"; issue: "20"; pages: "2492-505"
COMMENT     Sequence Label: pPA016 (delta 86-130 Parkin)
FEATURES             Location/Qualifiers
     source          1..6229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2279..2301)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2446..2464)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2532..2636
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2637..2705,2706..3497)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2855..2874)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3668..4256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4157..4176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4410..4427
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4500..4577
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4578..5660
                     /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(4597..4616)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5709..5714,5715..5732,5733..5739)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5747..5763
                     /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     5771..5787
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5771..5787
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5783..6229,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5803..5820)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5803..5819)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5812..5834)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc atgatcgttt
      961 tcgttcgttt caactcttct tacggtttcc cggttgaagt tgactctgac acctctatct
     1021 tccagctgaa agaagttgtt gcgaaacgtc agggtgttcc ggcggaccag ctgcgtgtta
     1081 tcttcgcggg taaagaactg cagaaccacc tgaccgttca gaactgcgac ctggaacagc
     1141 agtctatcgt tcacatcgtt cagcgtccgc agcgtaaatc tcacgaaacc aacgcgtctg
     1201 gtggttctga agcggcgcgt ggtccggcgg cgaaaccgac ctaccactct ttcttcgttt
     1261 actgcaaagg tccgtgccac aaagttcagc cgggtaaact gcgtgttcag tgcggtacct
     1321 gccgtcaggc gaccctgacc ctggcgcagg gtccgtcttg ctgggacgac gttctgatcc
     1381 cgaaccgtat gtctggtgaa tgccagtctc cggactgccc gggtacccgt gcggaattct
     1441 tcttcaaatg cggtgcgcac ccgacctctg acaaagacac ctctgttgcg ctgaacctga
     1501 tcaccaacaa ctctcgttct atcccgtgca tcgcgtgcac cgacgttcgt aacccggttc
     1561 tggttttcca gtgcaaccac cgtcacgtta tctgcctgga ctgcttccac ctgtactgcg
     1621 ttacccgtct gaacgaccgt cagttcgttc acgacgcgca gctgggttac tctctgccgt
     1681 gcgttgcggg ttgcccgaac tctctgatca aagaactgca ccacttccgt atcctgggtg
     1741 aagaacagta caaccgttac cagcagtacg gtgcggaaga atgcgttctg cagatgggtg
     1801 gtgttctgtg cccgcgtccg ggttgcggtg cgggtctgct gccggaacag ggtcagcgta
     1861 aagttacctg cgaaggtggt aacggtctgg gttgcggttt cgttttctgc cgtgactgca
     1921 aagaagcgta ccacgaaggt gaatgcgact ctatgttcga agcgtctggt gcgacctctc
     1981 aggcgtaccg tgttgaccag cgtgcggcgg aacaggcgcg ttgggaagaa gcgtctaaag
     2041 aaaccatcaa aaaaaccacc aaaccgtgcc cgcgttgcaa cgttccgatc gaaaaaaacg
     2101 gtggttgcat gcacatgaaa tgcccgcagc cgcagtgcaa actggaatgg tgctggaact
     2161 gcggttgcga atggaaccgt gcgtgcatgg gtgaccactg gttcgacgtt tagctcgagc
     2221 ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     2281 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     2341 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     2401 agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc
     2461 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     2521 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2581 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2641 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2701 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2761 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2821 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     2881 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2941 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3001 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3061 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3121 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3181 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3241 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3301 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3361 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3421 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3481 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3541 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3601 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3661 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3721 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3781 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3841 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3901 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3961 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4021 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4081 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4141 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4201 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4261 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4321 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4381 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4441 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg
     4501 acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     4561 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     4621 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     4681 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     4741 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     4801 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     4861 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     4921 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     4981 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     5041 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
     5101 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     5161 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     5221 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
     5281 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
     5341 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
     5401 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
     5461 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     5521 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac cacgctagcg cccaatacgc
     5581 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     5641 gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca
     5701 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     5761 caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca
     5821 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     5881 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     5941 cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag
     6001 ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat
     6061 gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc
     6121 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag
     6181 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatt
//
