LOCUS       sequence-169516-        5595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of hPPT in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5595)
  AUTHORS   Lu JY, Hu J, Hofmann SL
  TITLE     Human recombinant palmitoyl-protein thioesterase-1 (PPT1) for 
            preclinical evaluation of enzyme replacement therapy for infantile 
            neuronal ceroid lipofuscinosis.
  JOURNAL   Mol Genet Metab. 2010 Apr;99(4):374-8. doi: 
            10.1016/j.ymgme.2009.12.002. Epub 2009 Dec 5.
  PUBMED    20036592
REFERENCE   2  (bases 1 to 5595)
  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"; doi: 
            "10.1016/j.ymgme.2009.12.002"; journalName: "Mol Genet Metab"; date:
            "2010-04"; volume: "99"; issue: "4"; pages: "374-8"
COMMENT     Sequence Label: pCMV5-hPPT1
FEATURES             Location/Qualifiers
     source          1..5595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    995..1617
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1194..1213)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1641..1661)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1646..1975
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1826..1961
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2013..2032)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2014..2032)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2046..2062)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2046..2062)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2059..2081)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2070..2086
                     /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(2094..2100,2101..2118,2119..2124))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2139..2160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2277..2294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2448..3036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2528..2547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3207..3998,3999..4067))
                     /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     3830..3849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4068..4172)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4199..4654
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4286..4305)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4496..4517
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4780..4802
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4794..4811
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4795..4811
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4972..5351
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5352..5555
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5506..5526
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atggcgtcgc ccggctgcct gtggctcttg gctgtggctc tcctgccatg gacctgcgct
       61 tctcgggcgc tgcagcatct ggacccgccg gcgccgctgc cgttggtgat ctggcatggg
      121 atgggagaca gctgttgcaa tcccttaagc atgggtgcta ttaaaaaaat ggtggagaag
      181 aaaatacctg gaatttacgt cttatcttta gagattggga agaccctgat ggaggacgtg
      241 gagaacagct tcttcttgaa tgtcaattcc caagtaacaa cagtgtgtca ggcacttgct
      301 aaggatccta aattgcagca aggctacaat gctatgggat tctcccaggg aggccaattt
      361 ctgagggcag tggctcagag atgcccttca cctcccatga tcaatctgat ctcggttggg
      421 ggacaacatc aaggtgtttt tggactccct cgatgcccag gagagagctc tcacatctgt
      481 gacttcatcc gaaaaacact gaatgctggg gcgtactcca aagttgttca ggaacgcctc
      541 gtgcaagccg aatactggca tgaccccata aaggaggatg tgtatcgcaa ccacagcatc
      601 ttcttggcag atataaatca ggagcggggt atcaatgagt cctacaagaa aaacctgatg
      661 gccctgaaga agtttgtgat ggtgaaattc ctcaatgatt ccattgtgga ccctgtagat
      721 tcggagtggt ttggatttta cagaagtggc caagccaagg aaaccattcc cttacaggag
      781 acctccctgt acacacagga ccgcctgggg ctaaaggaaa tggacaatgc aggacagcta
      841 gtgtttctgg ctacagaagg ggaccatctt cagttgtctg aagaatggtt ttatgcccac
      901 atcataccat tccttggatg aaacccgtat agttcacaat agagctcagg gagccccaag
      961 cttgcatgcc tgcaggtcga ctctagagga tcccgggtgg catccctgtg acccctcccc
     1021 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     1081 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     1141 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     1201 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     1261 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     1321 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     1381 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     1441 ggcgtgaacc actgctccct tccctgtcct tctgatttta aaataactat accagcagga
     1501 ggacgtccag acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg
     1561 cttgcttggc actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg
     1621 tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     1681 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     1741 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1801 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     1861 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     1921 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     1981 cctcgaggaa ctgaaaaacc agaaagttaa ttccctatag tgagtcgtat taaattcgta
     2041 atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2101 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2161 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2221 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2281 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2341 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2401 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2461 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2521 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2581 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2641 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2701 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2761 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2821 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2881 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2941 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3001 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3061 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3121 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3181 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3241 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3301 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3361 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3421 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3481 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3541 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3601 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3661 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3721 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3781 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3841 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3901 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3961 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4021 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4081 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4141 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4201 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4261 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4321 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4381 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4441 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4501 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4561 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4621 gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact
     4681 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     4741 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     4801 cgacggccag tgccaagctg atctatacat tgaatcaata ttggcaatta gccatattag
     4861 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
     4921 atcataatat gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat
     4981 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     5041 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     5101 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     5161 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     5221 atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     5281 cccagtacat gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg
     5341 ctattaccat ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact
     5401 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     5461 atcaacggga ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta
     5521 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagaattcgt
     5581 ggtgacacag cgaag
//
