LOCUS       sequence-329853-        4959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for N. furzeri VPS35.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4959)
  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 4959)
  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-VPS35
FEATURES             Location/Qualifiers
     source          1..4959
                     /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(3084..3183)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3200..3219)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3201..3219)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3224..3240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3224..3240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3342..3361)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3353..4162
                     /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(3420..3439)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4309..4897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4798..4817
                     /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 acaaaatgat gccaaccaca cagcagtctc ctcaggatga
      721 gcaagagaag cttctagatg aagctgttca agctgtgaag gtccagtcct tccaaatgaa
      781 gcgatgcctg gacaaaaaca agctgatgga tgctttgaag catgcttcca atatgcttgg
      841 agagctgagg acttctatgc tttctccaaa aagttactat gagctctata tggccatctc
      901 cgatgaactt cactatcttg aggtttactt gactgatgag tttgcaaaag gacgcaaagt
      961 ggcagatctt tatgagctgg tccagtacgc tggcaacatt ataccccgac tctatctgct
     1021 gattacagtg ggtgtggtgt acgttcgctc tttccctcag tctcgcaagg acattttaaa
     1081 ggatctggtg gagatgtgtc gtggtgtcca gcacccactc agaggcctgt tcctcagaaa
     1141 ttacctgctg cagtgtaccc gcaacatcct gccagatgat ggcgagcagc cagagggaac
     1201 agaggagatg actggtgata tcaatgactc catcgacttc gttcttctta actttgcaga
     1261 aatgaacaaa ttatgggttc gaatgcagca tcaaggtcac agcagagaca gagagaagcg
     1321 agagaaggag cgccaggagc tcaggatttt agttggcacc aatctagttc gcttgagcca
     1381 attggagggt gtcaatgtgg acaagtacaa acagatagtt ttaccgggcg tgctggagca
     1441 ggttgtgaac tgtagagact cactggctca ggaatatctg atggagtgca taattcaggt
     1501 tttccctgat gagttccacc ttcaaacatt gaatcctttc ctgcgttctt gtgcggagct
     1561 gcatcaacac gtcaatgtga agaacatcat cattgccctc attgacagac tggctctctt
     1621 tgctcatcga gaggatggcc cgggtatccc agctgagatc aagttatttg acatcttctc
     1681 tcaacaagtg gccactgtca ttcaatctcg tcaagacatg ccttcagagg atgtggtttc
     1741 tctccaagtc tcgctcatca acttggccat gaagtgctac ccagaacgtg ttgactacgt
     1801 cgacaaggtc ctggagagca cagtggaaat cttcaataag cttaatttgg aacacattgc
     1861 gaccagcagt gccgtgtcta aagaactgac tcgacttttg aagatccctg tagacaccta
     1921 caacaacgtg ctgatggtgc tacagctcaa gcacttccca ccgctctttg aatactttga
     1981 ctttgagtcc cgcaagagca tgagctgcta tgtgctgagc aacacactcg acttcaacac
     2041 cgttattgtg gcacaagaac aggtggatgc catcttgagc ttggtgtcta cactgatcca
     2101 ggaccagcca gaccagccag ctgataatcc cgacccagag gactttgctg aggagcagag
     2161 ccttgtgggt cgtttcatcc atctttttca ttctgaagat cctgatcagc aatatcttat
     2221 actcaacaca gcccgaaaac attttggtgc aggcggaaat caaaggattc gttttacgtt
     2281 gccccccctg gtttttgctg cctaccagct ggccttcaga tacaaagaca attcctcgac
     2341 cgatgataag tgggagaaga agtgccagaa gatcttttct tttgcccacc agaccatcgg
     2401 tgccctcatc aaggctgagc tgtcagaact gcctctgcgg ctattcctgc agggggcgct
     2461 ggctgctggg gaaattggct ttgagaacca cgaaactgta gcgtatgagt ttatgtcaca
     2521 ggcattctcc ctgtatgagg atgagatcag tgactccaaa gcgcagctgg cagccatcac
     2581 attgatcatc ggcacctttg agcgaatgag atgtttcagc gaggagaatc acgagccttt
     2641 aaggactcag tgtgctctgg ctgcttccaa acttctgaag aagcctgacc agtgtcgagc
     2701 tgtcagcacg tgtgctcatc tgttctggtc gggtcgcagc actgacaaga atggtgaaga
     2761 gatccgtgac gggaagcgag tgatggagtg cctgaagaaa gccctgaaga tagccaacca
     2821 gtgtatggac tcatccctgc aggtccagct tttcattgag attctcaaca gatacatctg
     2881 cttctacgag agagaaaacg atgcggtgac ggtccaagtg ctgaaccaac tgattcaaaa
     2941 ggtcagagat gatcttccta atctggaagc aagtgaagaa actgagcaga tcaacaaaca
     3001 cttccacaac acactggagc accttcgtct gcagagagaa tctcccgagt ctgaaggtcc
     3061 tgcgtacgag ggtctggtcc ttgacccagc tttcttgtac aaagttggca ttataagaaa
     3121 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     3181 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     3241 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     3301 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     3361 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     3421 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     3481 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3541 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3601 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3661 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3721 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3781 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3841 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3901 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3961 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     4021 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     4081 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     4141 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     4201 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     4261 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4321 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4381 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4441 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4501 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4561 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4621 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4681 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4741 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4801 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4861 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4921 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
