LOCUS       sequence-172733-        6157 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 6157)
  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 6157)
  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-Myc-Parkin
FEATURES             Location/Qualifiers
     source          1..6157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      963..972
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             972..1001
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2457..2466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2470..2604
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2507..2526)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2561..2580
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2673..3030
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2673..2864
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2696..2716)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2881..3016
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3050..3067)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3050..3066)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3059..3081)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3279..3734
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3366..3385)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3576..3597
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3751..3770)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3870..3892
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3930..3948)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4016..4120
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4121..4189,4190..4981)
                     /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(4339..4358)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5152..5740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5641..5660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5894..5911
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6028..6049
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6064..6069,6070..6087,6088..6094)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6102..6118
                     /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     6107..6129
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6126..6142
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6126..6142
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcccgaac cgacagtcgg tctcttcacc aaggccattc
      961 gcgccaccat ggagcaaaag ctcatttctg aggaagatct caatggtggt ggtggtgggt
     1021 cgaccatgat agtgtttgtc aggttcaact ccagccatgg tttcccagtg gaggtcgatt
     1081 ctgacaccag catcttccag ctcaaggagg tggttgctaa gcgacagggg gttccggctg
     1141 accagttgcg tgtgattttc gcagggaagg agctgaggaa tgactggact gtgcagaatt
     1201 gtgacctgga tcagcagagc attgttcaca ttgtgcagag accgtggaga aaaggtcaag
     1261 aaatgaatgc aactggaggc gacgacccca gaaacgcggc gggaggctgt gagcgggagc
     1321 cccagagctt gactcgggtg gacctcagca gctcagtcct cccaggagac tctgtggggc
     1381 tggctgtcat tctgcacact gacagcagga aggactcacc accagctgga agtccagcag
     1441 gtagatcaat ctacaacagc ttttatgtgt attgcaaagg cccctgtcaa agagtgcagc
     1501 cgggaaaact cagggtacag tgcagcacct gcaggcaggc aacgctcacc ttgacccagg
     1561 gtccatcttg ctgggatgat gttttaattc caaaccggat gagtggtgaa tgccaatccc
     1621 cacactgccc tgggactagt gcagaatttt tctttaaatg tggagcacac cccacctctg
     1681 acaaggaaac atcagtagct ttgcacctga tcgcaacaaa tagtcggaac atcacttgca
     1741 ttacgtgcac agacgtcagg agccccgtcc tggttttcca gtgcaactcc cgccacgtga
     1801 tttgcttaga ctgtttccac ttatactgtg tgacaagact caatgatcgg cagtttgttc
     1861 acgaccctca acttggctac tccctgcctt gtgtggctgg ctgtcccaac tccttgatta
     1921 aagagctcca tcacttcagg attctgggag aagagcagta caaccggtac cagcagtatg
     1981 gtgcagagga gtgtgtcctg cagatggggg gcgtgttatg cccccgccct ggctgtggag
     2041 cggggctgct gccggagcct gaccagagga aagtcacctg cgaagggggc aatggcctgg
     2101 gctgtgggtt tgccttctgc cgggaatgta aagaagcgta ccatgaaggg gagtgcagtg
     2161 ccgtatttga agcctcagga acaactactc aggcctacag agtcgatgaa agagccgccg
     2221 agcaggctcg ttgggaagca gcctccaaag aaaccatcaa gaaaaccacc aagccctgtc
     2281 cccgctgcca tgtaccagtg gaaaaaaatg gaggctgcat gcacatgaag tgtccgcagc
     2341 cccagtgcag gctcgagtgg tgctggaact gtggctgcga gtggaaccgc gtctgcatgg
     2401 gggaccactg gttcgacgtg taggcggccg ctaagtaagt aaggatcccc agcttggccg
     2461 ccatggccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2521 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2581 aatgtatctt atcatgtctg gatcgggaat taattcggcg cagcaccatg gcctgaaata
     2641 acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat
     2701 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2761 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2821 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2881 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2941 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3001 ggcttttttg gaggcctagg cttttgcaaa aagctgttaa cagcttggca ctggccgtcg
     3061 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3121 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3181 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     3241 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     3301 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3361 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3421 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3481 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3541 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3601 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     3661 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3721 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3781 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3841 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3901 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3961 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4021 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4081 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4141 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4201 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4261 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4321 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4381 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4441 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4501 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4561 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4621 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4681 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4741 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4801 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4861 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4921 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4981 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5041 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5101 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5161 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5221 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5281 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5341 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5401 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5461 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5521 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5581 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5641 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5701 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5761 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5821 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5881 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     5941 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6001 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6061 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     6121 tcacacagga aacagctatg acatgattac gaattaa
//
