LOCUS       sequence-362509-        7918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SNAPtag-PTS1 in mammalian cells. SNAPtag targets to 
            peroxisomes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7918)
  AUTHORS   Bui H, Andersson S, Sola-Carvajal A, De Marchi T, Vahakangas E, 
            Holopainen M, House AH, Rovenko BM, Englund JI, Kasper M, 
            Kuuluvainen E, Kakela R, Hietakangas V, Nimeus E, Katajisto P
  TITLE     Glucose-6-phosphate-dehydrogenase on old peroxisomes maintains 
            self-renewal of epithelial stem cells after asymmetric cell 
            division.
  JOURNAL   Nat Commun. 2025 Apr 26;16(1):3932. doi: 10.1038/s41467-025-58752-z.
  PUBMED    40287409
REFERENCE   2  (bases 1 to 7918)
  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 
            Commun."; date: "2025-04-26"; volume: "16(1):3932. doi"; pages: " 
            10.1038/s41467-025-58752-z"
COMMENT     Sequence Label: SNAPtag-PTS1
FEATURES             Location/Qualifiers
     source          1..7918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        131..334
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     285..305
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             366..911
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for mammalian expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     misc_feature    1001..1118
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1167..1677
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1596..1614
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1699..2298
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1699..1718)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2195..2215
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2426..2659
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2731..2852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2768..2787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2822..2841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2892..3027
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3047..3066)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3048..3066)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3076..3093)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3076..3092)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3085..3107)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3234..3689
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3321..3340)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3531..3552
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3715..3819
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3820..3888,3889..4680)
                     /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(4038..4057)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4851..5439
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5340..5359
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5593..5610
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5727..5748
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5763..5768,5769..5786,5787..5793)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5801..5817
                     /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)."
     primer_bind     5806..5828
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5825..5841
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5825..5841
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5860..5880
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5862..5880
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5908..6134
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6135..6315
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6362..6487
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6980..7213
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7398..7442
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     primer_bind     7740..7760
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcgccatgga caaagattgc gaaatgaaac gtaccaccct ggatagccct ctgggcaagc
      421 tggaactgtc tgggtgcgaa cagggcctgc acgagatcaa gctgctgggc aaaggaacat
      481 ctgccgccga cgccgtggaa gtgcctgccc cagccgccgt gctgggcgga ccagagccac
      541 tgatgcaggc caccgcctgg ctcaacgcct actttcacca gcctgaggcc atcgaggagt
      601 tccctgtgcc agccctgcac cacccagtgt tccagcagga gagctttacc cgccaggtgc
      661 tgtggaaact gctgaaagtg gtgaagttcg gagaggtcat cagctaccag cagctggccg
      721 ccctggccgg caatcccgcc gccaccgccg ccgtgaaaac cgccctgagc ggaaatcccg
      781 tgcccattct gatcccctgc caccgggtgg tgtctagctc tggcgccgtg gggggctacg
      841 agggcgggct cgccgtgaaa gagtggctgc tggcccacga gggccacaga ctgggcaagc
      901 ctgggctggg tcctgcaggt ataggcgcgc caggatccac gcgtaagagc aagctgtgag
      961 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     1021 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     1081 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     1141 gagatccact ttggccgcgg ctcgaggggg ttggggttgc gccttttcca aggcagccct
     1201 gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag cggcgccgac
     1261 cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt cgccgctacc
     1321 cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg
     1381 ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac cctcgcagac
     1441 ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc
     1501 tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga ggcggggtgt
     1561 ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg caagcctccg
     1621 gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagggg
     1681 gatccaccgg agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     1741 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     1801 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     1861 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     1921 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     1981 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     2041 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     2101 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     2161 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     2221 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     2281 cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca
     2341 cgaccccatg catcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     2401 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     2461 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     2521 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     2581 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     2641 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     2701 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     2761 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2821 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     2881 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2941 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3001 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     3061 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     3121 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     3181 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     3241 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3301 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3361 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3421 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3481 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3541 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3601 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3661 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     3721 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3781 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3841 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3901 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3961 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4021 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4081 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4141 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4201 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4261 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4321 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4381 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4441 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4501 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4561 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4621 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4681 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4741 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4801 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4861 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4921 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4981 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5041 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5101 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5161 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5221 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5281 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5341 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5401 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5461 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5521 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5581 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     5641 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     5701 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     5761 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     5821 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     5881 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     5941 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
     6001 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     6061 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     6121 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     6181 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     6241 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     6301 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     6361 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     6421 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     6481 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     6541 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     6601 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     6661 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     6721 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     6781 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     6841 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     6901 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     6961 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     7021 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     7081 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     7141 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     7201 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     7261 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     7321 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     7381 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     7441 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     7501 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     7561 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
     7621 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
     7681 tgaacggatc tcgacggtat cgatcacgag actagcctcg agcggccgcc cccttcaccg
     7741 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
     7801 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
     7861 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatca
//
