LOCUS       sequence-396306-        8130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PB plasmid expressing RPS27A with its 3'UTR tagged to PP7 stem 
            loops.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8130)
  AUTHORS   De Pace R, Ghosh S, Ryan VH, Sohn M, Jarnik M, Rezvan Sangsari P, 
            Morgan NY, Dale RK, Ward ME, Bonifacino JS
  TITLE     Messenger RNA transport on lysosomal vesicles maintains axonal 
            mitochondrial homeostasis and prevents axonal degeneration.
  JOURNAL   Nat Neurosci. 2024 Apr 10. doi: 10.1038/s41593-024-01619-1.
  PUBMED    38600167
REFERENCE   2  (bases 1 to 8130)
  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: "Nat 
            Neurosci. 2024 Apr 10. doi: 10.1038/s41593-024-01619-1."
COMMENT     Sequence Label: PB-Ef1a-RPS27A-UTR-PP7-WPRE
FEATURES             Location/Qualifiers
     source          1..8130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     377..399
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     391..408
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     392..408
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   538..600
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        885..1096
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          1097..2035
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1992..2012
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2057..2284
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     protein_bind    complement(4303..4336)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4392..4980
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4445..4465)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4863..4874)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4983..4999)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4984..5000)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     repeat_region   5403..5437
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5928..5944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5928..5944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5941..5963)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5952..5968
                     /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)."
     promoter        complement(join(5976..5982,5983..6000,6001..6006))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6021..6042
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6159..6176)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6378..6966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6458..6477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7137..7928,7929..7997))
                     /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     7760..7779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7998..8102)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
      301 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
      361 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcctcg
      421 ttcattcacg tttttgaacc cgtggaggac gggcagactc gcggtgcaaa tgtgttttac
      481 agcgtgatgg agcagatgaa gatgctcgac acgctgcaga acacgcagct agattaaccc
      541 tagaaagata atcatattgt gacgtacgtt aaagataatc atgtgtaaaa ttgacgcatg
      601 tgttttatcg gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat
      661 tatatttaca cttacatact aataataaat tcaacaaaca atttatttat gtttatttat
      721 ttattaaaaa aaacaaaaac tcaaaatttc ttctataaag taacaaaact tttatgggcc
      781 cgctctagag aacctgaatg cagtctctcg gagggcgaag aatcctgcag gctcgaggcg
      841 gccgcgtgga taaccgtatt accgccatgc atgtgcccgt cagtgggcag agcgcacatc
      901 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
      961 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     1021 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     1081 tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg
     1141 ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc
     1201 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
     1261 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
     1321 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
     1381 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
     1441 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
     1501 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
     1561 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
     1621 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
     1681 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
     1741 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     1801 ccgggcgccg tccaggcacc tcgattagtt ctcgcgcttt tggagtacgt cgtctttagg
     1861 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     1921 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     1981 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     2041 gtgagctagc gccaccatgc agattttcgt gaaaaccctt acggggaaga ccatcaccct
     2101 cgaggttgaa ccctcggata cgatagaaaa tgtaaaggcc aagatccagg ataaggaagg
     2161 aattcctcct gatcagcaga gactgatctt tgctggcaag cagctggaag atggacgtac
     2221 tttgtctgac tacaatattc aaaaggagtc tactcttcat cttgtgttga gacttcgtgg
     2281 tggtgctaag aaaaggaaga agaagtctta caccactccc aagaagaata agcacaagag
     2341 aaagaaggtt aagctggctg tcctgaaata ttataaggtg gatgagaatg gcaaaattag
     2401 tcgccttcgt cgagagtgcc cttctgatga atgtggtgct ggggtgttta tggcaagtca
     2461 ctttgacaga cattattgtg gcaaatgttg tctgacttac tgtttcaaca aaccagaaga
     2521 caagtaactg tatgagttaa taaaagacat gaactaacat ttattgttgg gttttattgc
     2581 agtaaaaaga atggttttta agcaccaaat tgatggtcac accatttcct tttagtagtg
     2641 ctactgctat cgctgtgtga atgttgcctc tggggattat gtgacccagt ggttctgtat
     2701 acctgccagg tgccaaccac ttgtaaaggt cttgatattt tcaattctta gactacctat
     2761 actttggcag aagttatatt taatgtaagt tgtctaaata taatgtacag gtaccggatc
     2821 ctaaggtacc taattgccta gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga
     2881 ctctagaaac cagcagagca tatgggctcg ctggctgcag tattcccggg ttcattagat
     2941 cctaaggtac ctaattgcct agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg
     3001 actctagaaa ccagcagagc atatgggctc gctggctgca gtattcccgg gttcattaga
     3061 tcctaaggta cctaattgcc tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc
     3121 gactctagaa accagcagag catatgggct cgctggctgc agtattcccg ggttcattag
     3181 atcctaaggt acctaattgc ctagaaagga gcagacgata tggcgtcgct ccctgcaggt
     3241 cgactctaga aaccagcaga gcatatgggc tcgctggctg cagtattccc gggttcatta
     3301 gatcctaagg tacctaattg cctagaaagg agcagacgat atggcgtcgc tccctgcagg
     3361 tcgactctag aaaccagcag agcatatggg ctcgctggct gcagtattcc cgggttcatt
     3421 agatcctaag gtacctaatt gcctagaaag gagcagacga tatggcgtcg ctccctgcag
     3481 gtcgactcta gaaaccagca gagcatatgg gctcgctggc tgcagtattc ccgggttcat
     3541 tagatcctaa ggtacctaat tgcctagaaa ggagcagacg atatggcgtc gctccctgca
     3601 ggtcgactct agaaaccagc agagcatatg ggctcgctgg ctgcagtatt cccgggttca
     3661 ttagatccta aggtacctaa ttgcctagaa aggagcagac gatatggcgt cgctccctgc
     3721 aggtcgactc tagaaaccag cagagcatat gggctcgctg gctgcagtat tcccgggttc
     3781 attagatcct aaggtaccta attgcctaga aaggagcaga cgatatggcg tcgctccctg
     3841 caggtcgact ctagaaacca gcagagcata tgggctcgct ggctgcagta ttcccgggtt
     3901 cattagatcc taaggtacct aattgcctag aaaggagcag acgatatggc gtcgctccct
     3961 gcaggtcgac tctagaaacc agcagagcat atgggctcgc tggctgcagt attcccgggt
     4021 tcattagatc ctaaggtacc taattgccta gaaaggagca gacgatatgg cgtcgctccc
     4081 tgcaggtcga ctctagaaac cagcagagca tatgggctcg ctggctgcag tattcccggg
     4141 ttcattagat cctaaggtac ctaattgcct agaaaggagc agacgatatg gcgtcgctcc
     4201 ctgcaggtcg actctagaaa ccagcagagc atatgggctc gctggctgca gtattcccgg
     4261 gttcattaga tctcgcgaag ggcgaattcg tcgagggacc taataacttc gtatagcata
     4321 cattatacga agttatacat gtttaagggt tccggttcca ctaggtacaa ttcgatatca
     4381 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     4441 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     4501 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     4561 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     4621 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     4681 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     4741 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     4801 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     4861 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     4921 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     4981 atcgataccg tcgacctcga tcgagaccta gaaaaacatg gagcaatcac aagtagcaat
     5041 acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt
     5101 tttccagtca cacctcaggt acctttaagg attctataac aagaaaatat atatataata
     5161 agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt taaatcttaa
     5221 aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat agttcaaaat
     5281 cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg cgactgagat
     5341 gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat atttcaagaa
     5401 tgcatgcgtc aattttacgc agactatctt tctagggtta atctagctgc atcaggatca
     5461 tatcgtcggg tcttttttcc ggctcagtca tcgcccaagc tggcgctatc tgggcatcgg
     5521 ggaggaagaa gcccgtgcct tttcccgcga ggttgaagcg gcatggaaag agtttgccga
     5581 ggatgactgc tgctgcattg acgttgagcg aaaacgcacg tttaccatga tgattcggga
     5641 aggtgtggcc atgcacgcct ttaacggtga actgttcgtt caggccacct gggataccag
     5701 ttcgtcgcgg cttttccgga cacagttccg gatggtcagc ccgaagcgca tcagcaaccc
     5761 gaacaatacc ggcgacagcc ggaactgccg tgccggtgtg cagattaatg acagcggtgc
     5821 ggcgctggga tattacgtca gcgaggacgg gtatcctggc tggatgccgc agaaatggac
     5881 atggataccc cgtgagttac ccggcgggcg cgcttggcgt aatcatggtc atagctgttt
     5941 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6001 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6061 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6121 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6181 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6241 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6301 aaccgtaaaa aggccgcgtt gctggcgttg gatccggccg cgttgctggc gttggatccg
     6361 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6421 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6481 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6541 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6601 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6661 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6721 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6781 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6841 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6901 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6961 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7021 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7081 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7141 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7201 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7261 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7321 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7381 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7441 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7501 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7561 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7621 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7681 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7741 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7801 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7861 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7921 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7981 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8041 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8101 cgcacatttc cccgaaaagt gccacctgac
//
