LOCUS       sequence-138517-        6163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6163)
  AUTHORS   Zhang Y, Gao J, Chung KK, Huang H, Dawson VL, Dawson TM
  TITLE     Parkin functions as an E2-dependent ubiquitin- protein ligase and 
            promotes the degradation of the synaptic vesicle-associated protein,
            CDCrel-1.
  JOURNAL   Proc Natl Acad Sci U S A 2000 Nov 21;97(24):13354-9.
  PUBMED    11078524
REFERENCE   2  (bases 1 to 6163)
  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: "Proc Natl 
            Acad Sci U S A"; date: "2000-11-21"; volume: "97(24)"; pages: 
            "13354-9"
COMMENT     Sequence Label: pRK5-HA-Parkin
FEATURES             Location/Qualifiers
     source          1..6163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(91..96,97..114,115..121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /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     134..156
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     153..169
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     153..169
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        198..577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        578..781
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     732..752
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     778..802
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1012..1030
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1012..1029
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1043..1062
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1147..1156
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1159..1185
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2647..2656
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2660..2794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2697..2716)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2751..2770
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2863..3220
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2863..3054
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2886..2906)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3071..3206
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3240..3257)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3240..3256)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3249..3271)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3469..3924
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3556..3575)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3766..3787
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3941..3960)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4060..4082
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4120..4138)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4206..4310
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4311..4379,4380..5171)
                     /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     complement(4529..4548)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5342..5930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5831..5850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6084..6101
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa
      181 ttaattcgag ctcgcccgac attgattatt gactagttat taatagtaat caattacggg
      241 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      301 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      361 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      421 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      481 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      541 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      601 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      661 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      721 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      781 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
      841 aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa cgcggattcc
      901 ccgtgccaag agtgacgtaa gtaccgccta tagagtctat aggcccaccc ccttggcttc
      961 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg
     1021 acactataga ataacatcca ctttgccttt ctctccacag gtgtccactc ccaggtccaa
     1081 ctgcacctcg gttctatcga ttgaattccc gaaccgacag tcggtctctt caccaaggcc
     1141 attcgcgcca ccatgggcta cccctatgat gtgcctgact acgcagatct caatggtggt
     1201 ggtggtgggt cgaccatgat agtgtttgtc aggttcaact ccagccatgg tttcccagtg
     1261 gaggtcgatt ctgacaccag catcttccag ctcaaggagg tggttgctaa gcgacagggg
     1321 gttccggctg accagttgcg tgtgattttc gcagggaagg agctgaggaa tgactggact
     1381 gtgcagaatt gtgacctgga tcagcagagc attgttcaca ttgtgcagag accgtggaga
     1441 aaaggtcaag aaatgaatgc aactggaggc gacgacccca gaaacgcggc gggaggctgt
     1501 gagcgggagc cccagagctt gactcgggtg gacctcagca gctcagtcct cccaggagac
     1561 tctgtggggc tggctgtcat tctgcacact gacagcagga aggactcacc accagctgga
     1621 agtccagcag gtagatcaat ctacaacagc ttttatgtgt attgcaaagg cccctgtcaa
     1681 agagtgcagc cgggaaaact cagggtacag tgcagcacct gcaggcaggc aacgctcacc
     1741 ttgacccagg gtccatcttg ctgggatgat gttttaattc caaaccggat gagtggtgaa
     1801 tgccaatccc cacactgccc tgggactagt gcagaatttt tctttaaatg tggagcacac
     1861 cccacctctg acaaggaaac atcagtagct ttgcacctga tcgcaacaaa tagtcggaac
     1921 atcacttgca ttacgtgcac agacgtcagg agccccgtcc tggttttcca gtgcaactcc
     1981 cgccacgtga tttgcttaga ctgtttccac ttatactgtg tgacaagact caatgatcgg
     2041 cagtttgttc acgaccctca acttggctac tccctgcctt gtgtggctgg ctgtcccaac
     2101 tccttgatta aagagctcca tcacttcagg attctgggag aagagcagta caaccggtac
     2161 cagcagtatg gtgcagagga gtgtgtcctg cagatggggg gcgtgttatg cccccgccct
     2221 ggctgtggag cggggctgct gccggagcct gaccagagga aagtcacctg cgaagggggc
     2281 aatggcctgg gctgtgggtt tgccttctgc cgggaatgta aagaagcgta ccatgaaggg
     2341 gagtgcagtg ccgtatttga agcctcagga acaactactc aggcctacag agtcgatgaa
     2401 agagccgccg agcaggctcg ttgggaagca gcctccaaag aaaccatcaa gaaaaccacc
     2461 aagccctgtc cccgctgcca tgtaccagtg gaaaaaaatg gaggctgcat gcacatgaag
     2521 tgtccgcagc cccagtgcag gctcgagtgg tgctggaact gtggctgcga gtggaaccgc
     2581 gtctgcatgg gggaccactg gttcgacgtg taggcggccg ctaagtaagt aaggatcccc
     2641 agcttggccg ccatggccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     2701 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2761 caaactcatc aatgtatctt atcatgtctg gatcgggaat taattcggcg cagcaccatg
     2821 gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca
     2881 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2941 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3001 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3061 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3121 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3181 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctgttaa cagcttggca
     3241 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     3301 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     3361 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     3421 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     3481 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3541 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3601 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3661 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     3721 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3781 aaactggaac aacactcaac cctatctcgg gctattcttt taatttataa gggattttgc
     3841 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3901 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     3961 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4021 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4081 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4141 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4201 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4261 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4321 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4381 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4441 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4501 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4561 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4621 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4681 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4741 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4801 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4861 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4921 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4981 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5041 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5101 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5161 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5221 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5281 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5341 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5401 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5461 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5521 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5581 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5641 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5701 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5761 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5821 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5881 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5941 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6001 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6061 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     6121 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atg
//
