LOCUS       Exported                5822 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCMVTNT PINK1 C-myc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5822)
  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 5822)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5822
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             1119..2861
                     /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             2880..2909
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    2936..3057
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2947..2966)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3001..3020
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3238..3693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3325..3344)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3535..3556
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3760..3779)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3879..3901
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3939..3957)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4025..4129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4130..4990
                     /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(4348..4367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5161..5749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5650..5669
                     /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 actcgagaat tcggtaccat ggcggtgcga caggcgctgg
     1141 gccgcggcct gcagctgggt cgagcgctgc tgctgcgctt cacgggcaag cccggccggg
     1201 cctacggctt ggggcggccg ggcccggcgg cgggctgtgt ccgcggggag cgtccaggct
     1261 gggccgcagg accgggcgcg gagcctcgca gggtcgggct cgggctccct aaccgtctcc
     1321 gcttcttccg ccagtcggtg gccgggctgg cggcgcggtt gcagcggcag ttcgtggtgc
     1381 gggcctgggg ctgcgcgggc ccttgcggcc gggcagtctt tctggccttc gggctagggc
     1441 tgggcctcat cgaggaaaaa caggcggaga gccggcgggc ggtctcggcc tgtcaggaga
     1501 tccaggcaat ttttacccag aaaagcaagc cggggcctga cccgttggac acgagacgct
     1561 tgcagggctt tcggctggag gagtatctga tagggcagtc cattggtaag ggctgcagtg
     1621 ctgctgtgta tgaagccacc atgcctacat tgccccagaa cctggaggtg acaaagagca
     1681 ccgggttgct tccagggaga ggcccaggta ccagtgcacc aggagaaggg caggagcgag
     1741 ctccgggggc ccctgccttc cccttggcca tcaagatgat gtggaacatc tcggcaggtt
     1801 cctccagcga agccatcttg aacacaatga gccaggagct ggtcccagcg agccgagtgg
     1861 ccttggctgg ggagtatgga gcagtcactt acagaaaatc caagagaggt cccaagcaac
     1921 tagcccctca ccccaacatc atccgggttc tccgcgcctt cacctcttcc gtgccgctgc
     1981 tgccaggggc cctggtcgac taccctgatg tgctgccctc acgcctccac cctgaaggcc
     2041 tgggccatgg ccggacgctg ttcctcgtta tgaagaacta tccctgtacc ctgcgccagt
     2101 acctttgtgt gaacacaccc agcccccgcc tcgccgccat gatgctgctg cagctgctgg
     2161 aaggcgtgga ccatctggtt caacagggca tcgcgcacag agacctgaaa tccgacaaca
     2221 tccttgtgga gctggaccca gacggctgcc cctggctggt gatcgcagat tttggctgct
     2281 gcctggctga tgagagcatc ggcctgcagt tgcccttcag cagctggtac gtggatcggg
     2341 gcggaaacgg ctgtctgatg gccccagagg tgtccacggc ccgtcctggc cccagggcag
     2401 tgattgacta cagcaaggct gatgcctggg cagtgggagc catcgcctat gaaatcttcg
     2461 ggcttgtcaa tcccttctac ggccagggca aggcccacct tgaaagccgc agctaccaag
     2521 aggctcagct acctgcactg cccgagtcag tgcctccaga cgtgagacag ttggtgaggg
     2581 cactgctcca gcgagaggcc agcaagagac catctgcccg agtagccgca aatgtgcttc
     2641 atctaagcct ctggggtgaa catattctag ccctgaagaa tctgaagtta gacaagatgg
     2701 ttggctggct cctccaacaa tcggccgcca ctttgttggc caacaggctc acagagaagt
     2761 gttgtgtgga aacaaaaatg aagatgctct ttctggctaa cctggagtgt gaaacgctct
     2821 gccaggcagc cctcctcctc tgctcatgga gggcagccct gaagctcgac gtcgacccgg
     2881 aacaaaaact tatttctgaa gaagatctgt aggcggccgc ttcgagcaga catgataaga
     2941 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3001 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3061 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     3121 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgggctg gcgtaatagc
     3181 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
     3241 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3301 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3361 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3421 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3481 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3541 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3601 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3661 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     3721 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     3781 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3841 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3901 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3961 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4021 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4081 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4141 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4201 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4261 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4321 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4381 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4441 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4501 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4561 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4621 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4681 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4741 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4801 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4861 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4921 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4981 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5041 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5101 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5161 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5221 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5281 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5341 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5401 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5461 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5521 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5581 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5641 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5701 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5761 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat
     5821 ct
//
