LOCUS       sequence-222654-        5816 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 5816)
  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 5816)
  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 C-myc
FEATURES             Location/Qualifiers
     source          1..5816
                     /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             1023..2765
                     /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"
     CDS             2778..2807
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    2834..2955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2845..2864)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2899..2918
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3136..3591
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3223..3242)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3433..3454
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3658..3677)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3777..3799
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3837..3855)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3923..4027
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4028..4096,4097..4888)
                     /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(4246..4265)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5059..5647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5548..5567
                     /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 gaattcggta
     1021 ccatggcggt gcgacaggcg ctgggccgcg gcctgcagct gggtcgagcg ctgctgctgc
     1081 gcttcacggg caagcccggc cgggcctacg gcttggggcg gccgggcccg gcggcgggct
     1141 gtgtccgcgg ggagcgtcca ggctgggccg caggaccggg cgcggagcct cgcagggtcg
     1201 ggctcgggct ccctaaccgt ctccgcttct tccgccagtc ggtggccggg ctggcggcgc
     1261 ggttgcagcg gcagttcgtg gtgcgggcct ggggctgcgc gggcccttgc ggccgggcag
     1321 tctttctggc cttcgggcta gggctgggcc tcatcgagga aaaacaggcg gagagccggc
     1381 gggcggtctc ggcctgtcag gagatccagg caatttttac ccagaaaagc aagccggggc
     1441 ctgacccgtt ggacacgaga cgcttgcagg gctttcggct ggaggagtat ctgatagggc
     1501 agtccattgg taagggctgc agtgctgctg tgtatgaagc caccatgcct acattgcccc
     1561 agaacctgga ggtgacaaag agcaccgggt tgcttccagg gagaggccca ggtaccagtg
     1621 caccaggaga agggcaggag cgagctccgg gggcccctgc cttccccttg gccatcaaga
     1681 tgatgtggaa catctcggca ggttcctcca gcgaagccat cttgaacaca atgagccagg
     1741 agctggtccc agcgagccga gtggccttgg ctggggagta tggagcagtc acttacagaa
     1801 aatccaagag aggtcccaag caactagccc ctcaccccaa catcatccgg gttctccgcg
     1861 ccttcacctc ttccgtgccg ctgctgccag gggccctggt cgactaccct gatgtgctgc
     1921 cctcacgcct ccaccctgaa ggcctgggcc atggccggac gctgttcctc gttatgaaga
     1981 actatccctg taccctgcgc cagtaccttt gtgtgaacac acccagcccc cgcctcgccg
     2041 ccatgatgct gctgcagctg ctggaaggcg tggaccatct ggttcaacag ggcatcgcgc
     2101 acagagacct gaaatccgac aacatccttg tggagctgga cccagacggc tgcccctggc
     2161 tggtgatcgc agattttggc tgctgcctgg ctgatgagag catcggcctg cagttgccct
     2221 tcagcagctg gtacgtggat cggggcggaa acggctgtct gatggcccca gaggtgtcca
     2281 cggcccgtcc tggccccagg gcagtgattg actacagcaa ggctgatgcc tgggcagtgg
     2341 gagccatcgc ctatgaaatc ttcgggcttg tcaatccctt ctacggccag ggcaaggccc
     2401 accttgaaag ccgcagctac caagaggctc agctacctgc actgcccgag tcagtgcctc
     2461 cagacgtgag acagttggtg agggcactgc tccagcgaga ggccagcaag agaccatctg
     2521 cccgagtagc cgcaaatgtg cttcatctaa gcctctgggg tgaacatatt ctagccctga
     2581 agaatctgaa gttagacaag atggttggct ggctcctcca acaatcggcc gccactttgt
     2641 tggccaacag gctcacagag aagtgttgtg tggaaacaaa aatgaagatg ctctttctgg
     2701 ctaacctgga gtgtgaaacg ctctgccagg cagccctcct cctctgctca tggagggcag
     2761 ccctgaagct cgacccggaa caaaaactta tttctgaaga agatctgtag gcggccgctt
     2821 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2881 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2941 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     3001 atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
     3061 ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3121 ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3181 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3241 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3301 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3361 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3421 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3481 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3541 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     3601 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     3661 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3721 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3781 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3841 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3901 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3961 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4021 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4081 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4141 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4201 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4261 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4321 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4381 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4441 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4501 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4561 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4621 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4681 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4741 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4801 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4861 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4921 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4981 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5041 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5101 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5161 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5221 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5281 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5341 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5401 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5461 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5521 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5581 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5641 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5701 tcacatggct cgacagatct tcaatattgg ccattagcca tattattcat tggttatata
     5761 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatat
//
