LOCUS       sequence-222763-        5815 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 5815)
  AUTHORS   Beilina A, Van Der Brug M, Ahmad R, Kesavapany S, Miller DW, Petsko 
            GA, Cookson MR
  TITLE     Mutations in PTEN-induced putative kinase 1 associated with 
            recessive parkinsonism have differential effects on protein 
            stability.
  JOURNAL   Proc Natl Acad Sci U S A. 2005 Apr 19. 102(16):5703-8.
  PUBMED    15824318
REFERENCE   2  (bases 1 to 5815)
  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. 2005 Apr 19. 102(16):5703-8."
COMMENT     Sequence Label: pCMVTNT PINK1 N-myc
FEATURES             Location/Qualifiers
     source          1..5815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     576..596
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     622..646
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          761..893
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     876..895
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        938..956
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        962..980
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     962..979
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1015..1044
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1057..2799
                     /codon_start=1
                     /product="human PTEN-induced putative kinase 1, a 
                     mitochondria serine/threonine protein kinase"
                     /label=PINK1
                     /translation="MAVRQALGRGLQLGRALLLRFTGKPGRAYGLGRPGPAAGCVRGER
                     PGWAAGPGAEPRRVGLGLPNRLRFFRQSVAGLAARLQRQFVVRAWGCAGPCGRAVFLAF
                     GLGLGLIEEKQAESRRAVSACQEIQAIFTQKSKPGPDPLDTRRLQGFRLEEYLIGQSIG
                     KGCSAAVYEATMPTLPQNLEVTKSTGLLPGRGPGTSAPGEGQERAPGAPAFPLAIKMMW
                     NISAGSSSEAILNTMSQELVPASRVALAGEYGAVTYRKSKRGPKQLAPHPNIIRVLRAF
                     TSSVPLLPGALVDYPDVLPSRLHPEGLGHGRTLFLVMKNYPCTLRQYLCVNTPSPRLAA
                     MMLLQLLEGVDHLVQQGIAHRDLKSDNILVELDPDGCPWLVIADFGCCLADESIGLQLP
                     FSSWYVDRGGNGCLMAPEVSTARPGPRAVIDYSKADAWAVGAIAYEIFGLVNPFYGQGK
                     AHLESRSYQEAQLPALPESVPPDVRQLVRALLQREASKRPSARVAANVLHLSLWGEHIL
                     ALKNLKLDKMVGWLLQQSAATLLANRLTEKCCVETKMKMLFLANLECETLCQAALLLCS
                     WRAAL"
     polyA_signal    2833..2954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2844..2863)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2898..2917
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3135..3590
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3222..3241)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3432..3453
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3657..3676)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3776..3798
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3836..3854)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3922..4026
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4027..4095,4096..4887)
                     /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(4245..4264)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5058..5646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5547..5566
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtacatttat attggctcat gtccaatatg accgccatgt tggcattgat tattgactag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
      601 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcacta gaagctttat
      661 tgcggtagtt tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc
      721 gaacttaagc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag
      781 acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt
      841 ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca
      901 ctcccagttc aattacagct cttaaggcta gagtacttaa tacgactcac tataggctag
      961 catttaggtg acactataga atacaagcta cttgttcttt ttgcactcga gatggaacaa
     1021 aaacttattt ctgaagaaga tctggaattc ggtaccatgg cggtgcgaca ggcgctgggc
     1081 cgcggcctgc agctgggtcg agcgctgctg ctgcgcttca cgggcaagcc cggccgggcc
     1141 tacggcttgg ggcggccggg cccggcggcg ggctgtgtcc gcggggagcg tccaggctgg
     1201 gccgcaggac cgggcgcgga gcctcgcagg gtcgggctcg ggctccctaa ccgtctccgc
     1261 ttcttccgcc agtcggtggc cgggctggcg gcgcggttgc agcggcagtt cgtggtgcgg
     1321 gcctggggct gcgcgggccc ttgcggccgg gcagtctttc tggccttcgg gctagggctg
     1381 ggcctcatcg aggaaaaaca ggcggagagc cggcgggcgg tctcggcctg tcaggagatc
     1441 caggcaattt ttacccagaa aagcaagccg gggcctgacc cgttggacac gagacgcttg
     1501 cagggctttc ggctggagga gtatctgata gggcagtcca ttggtaaggg ctgcagtgct
     1561 gctgtgtatg aagccaccat gcctacattg ccccagaacc tggaggtgac aaagagcacc
     1621 gggttgcttc cagggagagg cccaggtacc agtgcaccag gagaagggca ggagcgagct
     1681 ccgggggccc ctgccttccc cttggccatc aagatgatgt ggaacatctc ggcaggttcc
     1741 tccagcgaag ccatcttgaa cacaatgagc caggagctgg tcccagcgag ccgagtggcc
     1801 ttggctgggg agtatggagc agtcacttac agaaaatcca agagaggtcc caagcaacta
     1861 gcccctcacc ccaacatcat ccgggttctc cgcgccttca cctcttccgt gccgctgctg
     1921 ccaggggccc tggtcgacta ccctgatgtg ctgccctcac gcctccaccc tgaaggcctg
     1981 ggccatggcc ggacgctgtt cctcgttatg aagaactatc cctgtaccct gcgccagtac
     2041 ctttgtgtga acacacccag cccccgcctc gccgccatga tgctgctgca gctgctggaa
     2101 ggcgtggacc atctggttca acagggcatc gcgcacagag acctgaaatc cgacaacatc
     2161 cttgtggagc tggacccaga cggctgcccc tggctggtga tcgcagattt tggctgctgc
     2221 ctggctgatg agagcatcgg cctgcagttg cccttcagca gctggtacgt ggatcggggc
     2281 ggaaacggct gtctgatggc cccagaggtg tccacggccc gtcctggccc cagggcagtg
     2341 attgactaca gcaaggctga tgcctgggca gtgggagcca tcgcctatga aatcttcggg
     2401 cttgtcaatc ccttctacgg ccagggcaag gcccaccttg aaagccgcag ctaccaagag
     2461 gctcagctac ctgcactgcc cgagtcagtg cctccagacg tgagacagtt ggtgagggca
     2521 ctgctccagc gagaggccag caagagacca tctgcccgag tagccgcaaa tgtgcttcat
     2581 ctaagcctct ggggtgaaca tattctagcc ctgaagaatc tgaagttaga caagatggtt
     2641 ggctggctcc tccaacaatc ggccgccact ttgttggcca acaggctcac agagaagtgt
     2701 tgtgtggaaa caaaaatgaa gatgctcttt ctggctaacc tggagtgtga aacgctctgc
     2761 caggcagccc tcctcctctg ctcatggagg gcagccctgt agctcgagtg cggccgcttc
     2821 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2881 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2941 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     3001 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc
     3061 cgggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     3121 tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3181 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3241 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3301 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3361 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3421 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3481 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3541 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg
     3601 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     3661 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     3721 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     3781 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     3841 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     3901 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3961 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4021 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4081 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4141 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4201 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4261 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4321 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4381 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4441 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4501 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4561 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4621 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4681 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4741 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4801 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4861 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4921 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4981 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5041 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5101 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5161 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5221 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5281 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5341 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5401 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5461 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5521 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5581 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5641 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5701 cacatggctc gacagatctt caatattggc cattagccat attattcatt ggttatatag
     5761 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatat
//
