LOCUS       sequence-254046-        6727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses protein UbcH7 (C86K) fused to Bactrocera parkin with 
            residues 1-29, 117-160 and 412-496 deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6727)
  AUTHORS   Sauve V, Sung G, Soya N, Kozlov G, Blaimschein N, Miotto LS, Trempe 
            JF, Lukacs GL, Gehring K
  TITLE     Mechanism of parkin activation by phosphorylation.
  JOURNAL   Nat Struct Mol Biol. 2018 Jul;25(7):623-630. doi: 
            10.1038/s41594-018-0088-7. Epub 2018 Jul 2.
  PUBMED    29967542
REFERENCE   2  (bases 1 to 6727)
  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: 
            "10.1038/s41594-018-0088-7"; journalName: "Nat Struct Mol Biol"; 
            date: "2018-07"; volume: "25"; issue: "7"; pages: "623-630"
COMMENT     Sequence Label: pGEX-UbcH7(C86K)-10aa-linker-delta 1-29, 117-1...
FEATURES             Location/Qualifiers
     source          1..6727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     247..264
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        337..414
                     /gene="lacI"
                     /label=lacI promoter
     CDS             415..1497
                     /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(434..453)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1546..1551,1552..1569,1570..1576)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1584..1600
                     /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     1608..1624
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1608..1624
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1620..2147
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1640..1657)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1640..1656)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1649..1671)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2249..2254,2255..2270,2271..2277)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2285..2301
                     /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             2324..2977
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2935..2957
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2984..3007
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     regulatory      3015..3024
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4843..4865)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5010..5028)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5096..5200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5201..5269,5270..6061)
                     /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(5419..5438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6232..6727,1..93)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     join(6721..6727,1..13)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
       61 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      121 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
      181 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
      241 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
      301 acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg gcgcaaaacc
      361 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
      421 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
      481 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
      541 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
      601 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
      661 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
      721 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
      781 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
      841 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
      901 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
      961 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     1021 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     1081 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     1141 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     1201 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     1261 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     1321 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     1381 ctggtgaaaa gaaaaaccac gctagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     1441 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     1501 caacgcaatt aatgtaagtt agctcactca ttaggcaccc caggctttac actttatgct
     1561 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     1621 tgaccatgat tacggattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     1681 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     1741 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct
     1801 ttgcctggtt tccggcacca gaagcggtgc cggaaagctg gctggagtgc gatcttcctg
     1861 aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct
     1921 acaccaacgt gacctatccc attacggtca atccgccgtt tgttcccacg gagaatccga
     1981 cgggttgtta ctcgctcaca tttaatgttg atgaaagctg gctacaggaa ggccagacgc
     2041 gaattatttt tgatggcgtt ggaattagct tatcgactgc acggtgcacc aatgcttctg
     2101 gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat
     2161 tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg
     2221 gttctggcaa atattctgaa atgagctgtt gacaattaat catcggctcg tataatgtgt
     2281 ggaattgtga gcggataaca atttcacaca ggaaacagta ttcatgtccc ctatactagg
     2341 ttattggaaa attaagggcc ttgtgcaacc cactcgactt cttttggaat atcttgaaga
     2401 aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt
     2461 tgaattgggt ttggagtttc ccaatcttcc ttattatatt gatggtgatg ttaaattaac
     2521 acagtctatg gccatcatac gttatatagc tgacaagcac aacatgttgg gtggttgtcc
     2581 aaaagagcgt gcagagattt caatgcttga aggagcggtt ttggatatta gatacggtgt
     2641 ttcgagaatt gcatatagta aagactttga aactctcaaa gttgattttc ttagcaagct
     2701 acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt taaatggtga
     2761 tcatgtaacc catcctgact tcatgttgta tgacgctctt gatgttgttt tatacatgga
     2821 cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat
     2881 cccacaaatt gataagtact tgaaatccag caagtatata gcatggcctt tgcagggctg
     2941 gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg gatctggaag ttctgttcca
     3001 ggggcccctg ggatccatgg cggccagcag gaggctgatg aaggagcttg aagaaatccg
     3061 caaatgtggg atgaaaaact tccgtaacat ccaggttgat gaagctaatt tattgacttg
     3121 gcaagggctt attgttcctg acaaccctcc atatgataag ggagccttca gaatcgaaat
     3181 caactttcca gcagagtacc cattcaaacc accgaagatc acatttaaaa caaagatcta
     3241 tcacccaaac atcgacgaaa aggggcaggt caaactgcca gtaattagtg ccgaaaactg
     3301 gaagccagca accaaaaccg accaagtaat ccagtccctc atagcactgg tgaatgaccc
     3361 ccagcctgag cacccgcttc gggctgacct agctgaagaa tactctaagg accgtaaaaa
     3421 attctgtaag aatgctgaag agtttacaaa gaaatatggg gaaaagcgac ctgtggacgg
     3481 cggtgaaaac ctgtacttgg ggggatccct gagcatctac atcaagacga acacgggtcg
     3541 tacgctgagc gtaaacctgg agccacaatg ggacatcaag aacgtgaagg agatcgtggc
     3601 acctcagctg ggtctgcaac ctgaggaggt aaagatcatc ttcgccggca aggagctgag
     3661 cgacgcaact actatccagg agtgtgacct gggtcagcaa agcatcctgc atgcaatccg
     3721 ttctcgtcct caaccacagc gtcagcgttc tgaggaacgt atcaacatca ccgaggagga
     3781 ccgtcagcgt actaaagcac acttcttcgt ccactgcgca cagtgcaaca aactgtgcaa
     3841 aggcaaactg cgcgtgcgtt gctctctgtg taaaggcggt gctttcaccg tacaccgtga
     3901 cccggaatgt tgggatgacg ttctgaaacc gcgtcgtatc actggtcatt gcgaatccca
     3961 ggaaatcgcc tgcttcgaca acgaaaccgg tgacccgccg tttactgaat tctacttcaa
     4021 atgcggcgaa catgtgtccg gcggtgaaaa agacttcgcg gcgccgctga acctgattaa
     4081 aattaacatt aaagatgtcc cgtgcctggc ttgcacggaa gtgagcgaaa ccgtgctggt
     4141 gttcccgtgc gaaagcaaac acgtcacctg cctggaatgc ttcgaacagt actgccgctc
     4201 ccgcctgtcc gaacgtcagt tcatgccgca tccggatatt ggttacaccc tgccgtgtcc
     4261 ggctggttgt gaaaactcct tcattgaaga aattcaccac ttcaaactgc tgtcccgcga
     4321 agaatacgcg cgctaccagc gttttgcgac cgaagaatac gttctgcagg cgggcggtgt
     4381 tctgtgcccg cagccgggtt gtggtatggg tctgctggtt gaaccggaat gtaaaaaagt
     4441 tacctgccag aacggttgcg gttatgtttt ttgccgcaac tgcctgcagg gctatcacct
     4501 gggtgattgc ctgccggaat aaacttctac caatgcttcc ggctcttgcg aatattctgt
     4561 tgatccgaat cgcgcggctg aagctcgttg ggatgaagcg tctaaagtta ccattaaagt
     4621 tatgaccaaa ccgtgcccga aatgccgcac cccgaccgaa cgtgatggcg gcgctatgca
     4681 catggtttgt acccgcgccg gctgtggctt tgaatggtgt tggatttgtc agaccgaatg
     4741 gacccgtgat tgtatgggcg cccactggtt tggctaactc gagcggccgc atcgtgactg
     4801 actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4861 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     4921 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     4981 cggagtgtat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5041 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5101 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5161 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5221 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5281 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5341 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5401 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     5461 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5521 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5581 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5641 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5701 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     5761 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5821 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5881 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5941 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6001 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6061 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6121 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6181 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6241 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6301 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6361 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6421 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6481 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6541 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6601 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6661 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6721 gggaaac
//
