LOCUS       sequence-7606-ex        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        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     718..742
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1034..1052
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1058..1076
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1058..1075
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1111..1140
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1153..2895
                     /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    2929..3050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2940..2959)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2994..3013
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3231..3686
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3318..3337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3528..3549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3753..3772)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3872..3894
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3932..3950)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4018..4122
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4123..4191,4192..4983)
                     /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(4341..4360)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5154..5742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5643..5662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt acttaatacg actcactata ggctagcatt taggtgacac tatagaatac
     1081 aagctacttg ttctttttgc actcgagatg gaacaaaaac ttatttctga agaagatctg
     1141 gaattcggta ccatggcggt gcgacaggcg ctgggccgcg gcctgcagct gggtcgagcg
     1201 ctgctgctgc gcttcacggg caagcccggc cgggcctacg gcttggggcg gccgggcccg
     1261 gcggcgggct gtgtccgcgg ggagcgtcca ggctgggccg caggaccggg cgcggagcct
     1321 cgcagggtcg ggctcgggct ccctaaccgt ctccgcttct tccgccagtc ggtggccggg
     1381 ctggcggcgc ggttgcagcg gcagttcgtg gtgcgggcct ggggctgcgc gggcccttgc
     1441 ggccgggcag tctttctggc cttcgggcta gggctgggcc tcatcgagga aaaacaggcg
     1501 gagagccggc gggcggtctc ggcctgtcag gagatccagg caatttttac ccagaaaagc
     1561 aagccggggc ctgacccgtt ggacacgaga cgcttgcagg gctttcggct ggaggagtat
     1621 ctgatagggc agtccattgg taagggctgc agtgctgctg tgtatgaagc caccatgcct
     1681 acattgcccc agaacctgga ggtgacaaag agcaccgggt tgcttccagg gagaggccca
     1741 ggtaccagtg caccaggaga agggcaggag cgagctccgg gggcccctgc cttccccttg
     1801 gccatcaaga tgatgtggaa catctcggca ggttcctcca gcgaagccat cttgaacaca
     1861 atgagccagg agctggtccc agcgagccga gtggccttgg ctggggagta tggagcagtc
     1921 acttacagaa aatccaagag aggtcccaag caactagccc ctcaccccaa catcatccgg
     1981 gttctccgcg ccttcacctc ttccgtgccg ctgctgccag gggccctggt cgactaccct
     2041 gatgtgctgc cctcacgcct ccaccctgaa ggcctgggcc atggccggac gctgttcctc
     2101 gttatgaaga actatccctg taccctgcgc cagtaccttt gtgtgaacac acccagcccc
     2161 cgcctcgccg ccatgatgct gctgcagctg ctggaaggcg tggaccatct ggttcaacag
     2221 ggcatcgcgc acagagacct gaaatccgac aacatccttg tggagctgga cccagacggc
     2281 tgcccctggc tggtgatcgc agattttggc tgctgcctgg ctgatgagag catcggcctg
     2341 cagttgccct tcagcagctg gtacgtggat cggggcggaa acggctgtct gatggcccca
     2401 gaggtgtcca cggcccgtcc tggccccagg gcagtgattg actacagcaa ggctgatgcc
     2461 tgggcagtgg gagccatcgc ctatgaaatc ttcgggcttg tcaatccctt ctacggccag
     2521 ggcaaggccc accttgaaag ccgcagctac caagaggctc agctacctgc actgcccgag
     2581 tcagtgcctc cagacgtgag acagttggtg agggcactgc tccagcgaga ggccagcaag
     2641 agaccatctg cccgagtagc cgcaaatgtg cttcatctaa gcctctgggg tgaacatatt
     2701 ctagccctga agaatctgaa gttagacaag atggttggct ggctcctcca acaatcggcc
     2761 gccactttgt tggccaacag gctcacagag aagtgttgtg tggaaacaaa aatgaagatg
     2821 ctctttctgg ctaacctgga gtgtgaaacg ctctgccagg cagccctcct cctctgctca
     2881 tggagggcag ccctgtgact cgagtgcggc cgcttcgagc agacatgata agatacattg
     2941 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     3001 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     3061 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt
     3121 aaaacctcta caaatgtggt aaaatcgata aggatccggg ctggcgtaat agcgaagagg
     3181 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     3241 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3301 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3361 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3421 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3481 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3541 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3601 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3661 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     3721 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3781 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3841 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3901 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3961 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4021 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4081 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4141 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4201 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4261 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4321 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4381 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4441 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4501 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4561 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4621 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4681 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4741 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4801 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4861 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4921 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4981 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5041 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5101 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5161 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5221 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5281 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5341 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5401 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5461 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5521 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5581 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5641 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5701 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5761 tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatct
//
