LOCUS       sequence-329581-        4866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for N. furzeri PNPT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4866)
  AUTHORS   Chen YR, Harel I, Singh PP, Ziv I, Moses E, Goshtchevsky U, Machado 
            BE, Brunet A, Jarosz DF
  TITLE     Tissue-specific landscape of protein aggregation and quality control
            in an aging vertebrate.
  JOURNAL   Dev Cell. 2024 Jul 22;59(14):1892-1911.e13. doi: 
            10.1016/j.devcel.2024.04.014. Epub 2024 May 28.
  PUBMED    38810654
REFERENCE   2  (bases 1 to 4866)
  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.devcel.2024.04.014"; journalName: "Dev Cell"; date: 
            "2024-07-22- 22"; volume: "59"; issue: "14"; pages: "1892-1911.e13"
COMMENT     Sequence Label: pDONR221-PNPT1
FEATURES             Location/Qualifiers
     source          1..4866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2991..3090)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3107..3126)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3108..3126)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3131..3147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3131..3147)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3249..3268)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3260..4069
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3327..3346)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4216..4804
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4705..4724
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc gaaggagata acaaaatgag gtgtttgctc cgactgggcc gctcgttcgg
      721 ccaccttcgg gtccggttat gtcaccagag tgtttacagc aggaacacaa acctccggag
      781 agtgggagtg aaccttggag agaaaaaact tgaaatctct tcggggaagt ttgcaaggtt
      841 tgctgatggt gctgctgtgg tacagctggg ggacacctct gtgatggtga ctgctgtgag
      901 caaaaccaaa ccttctccat ctcagttcat gccacttgtg gtggattaca gacagaaggc
      961 tgctgctgca ggtcgaatcc caaccaacta tttgaggaga gaactaggca ccactgataa
     1021 tgagatcctc accagcagac tcatcgatcg gtcgattaga cctcttttcc ctcctggcta
     1081 cttttatgac actcaggtcc tttgtaacct cctcgcagtt gatggtgtaa atgacccaga
     1141 cgtgctagct attaatgcag cttctgctgc tctggctctg tctgacatcc cttggaatgg
     1201 ccccataggt gctgtacgag ttggtatggt taatggagag ttactggtga acccgacaag
     1261 gacagagatg gcttccagca gtttgaacat gattgtggct ggggctccta gcagtcaagt
     1321 ggtgatgatt gaggcatcgg ctgagaacgt tctgcagcag gacttctgcc gtgcggtaaa
     1381 ggtgggagtc aaacacaccc agcagatcat actggccatc cagcagctgg tgcgggagca
     1441 gaaagtcatc aaacgcacgc cagtcagaat gttctcagtc ccgactgaca tggtggaaca
     1501 cgtccagaaa ttggtgtctg agaagatcca tgctgttttc acagattact cgcatgataa
     1561 gatttcgaga gacgaagcga ttaacaaaat acgactagaa actgaggaga actttaaaga
     1621 aaaattccct caggcggagc cctttgaagt cattgaatca ttcaatgctg tgaacaaaga
     1681 agttttccga aatttagttc tcagtgagta caagaggtgt gatggaaggg aactgacgtc
     1741 tttaaggaac atatcctgtg acattgacct cttcaagccg ctccatggct cagctttgtt
     1801 tcagagagga cagacacagg tgctctgcag tgttacattt gactccctgg aatcaagtgt
     1861 caaaacagac attataacca ctgctctcag tggcatcaaa gacaaaaact ttatgcttca
     1921 ttatgagttc cctccttatg caacaaacga gactgggaaa atgggaggtc tcaacaggag
     1981 agagcttggg catggagctc tggctgagaa ggctcttaga ccggtcattc ctacagattt
     2041 ccctttcacc atcagggtca cagcagaggt gctggagtcc aacggatcct cctcgatggc
     2101 ttcagcgtgt ggaggcagcc ttgctctaat ggatgcaggc gttcccatct cttcagctgt
     2161 agcaggcgtc gccgtcggcc tgatctcaaa ggctaatcca gagaaacccg gcgagatgga
     2221 ggactacaga ttactaacag atattttagg aatagaggat tacctgggag acatggactt
     2281 caaactggca ggaacaaaca agggtatcac tgcattacag gctgatgtga agattccagg
     2341 tttacctctt aaggtggtca tggaggcgat tcagcaggcc acagtggcaa agagagaaat
     2401 cttgggtatt atgaataaat gtatagcaaa gcccagaggg accaggaagg agaatggacc
     2461 tgttgttgag aatgtcaggg tacctctgtc tagaagagct cgcttcgttg gtccaggagg
     2521 atgtaacctc cgaaagcttc aagctcagac gggcgttaca ataagtcagg tggacgagga
     2581 gaccttttct gtgttcgctc ccacaccagg agccatgaat gaagcccaag agtttatcag
     2641 agaggtctgc aaagatgatc aagaacagca gctggagttt ggtgctgtct acactgccac
     2701 catcacagaa ataagggata ttggagtgat ggtgaaactt tatcccaaca tgagtccagt
     2761 gctactgcac aactcccagc tggaccataa acggatcaaa catccaagtg ctttgggctt
     2821 agaagtggga caggagatac aggtcaagta ctttggacgg gatccaacag atggaaggat
     2881 gaggctttcg cgaaaggtgc tccagtctcc tgctgcaact gtggttaaaa ccctgagtga
     2941 gaaacagagc atctctatgg gcaacagcaa caggaccgaa atggacgtag acccagcttt
     3001 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     3061 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     3121 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     3181 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     3241 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     3301 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     3361 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     3421 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     3481 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     3541 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     3601 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     3661 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     3721 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3781 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3841 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3901 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3961 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     4021 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     4081 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     4141 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     4201 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4261 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4321 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4381 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4441 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4501 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4561 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4621 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4681 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4741 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4801 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4861 catgtt
//
