LOCUS       sequence-329506-        4164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for N. furzeri EHD1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4164)
  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 4164)
  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-EHD1
FEATURES             Location/Qualifiers
     source          1..4164
                     /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(2289..2388)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2405..2424)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2406..2424)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2429..2445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2429..2445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2547..2566)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2558..3367
                     /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(2625..2644)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3514..4102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4003..4022
                     /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 gtttagaaaa accctcaaga aggacccgga
      721 gttgtttcaa aacgtgtcgg atgggttacg tcggctttac cgaaccaaac tgttcccact
      781 ggaggacacg tatcgattcc acgacttcca ctcccctgca ctcgaagatg ccgacttcga
      841 taacaagccc atggtacttt tggtaggcca gtactccacc ggtaaaacca cctttatccg
      901 acaccttatt gagcaggact tccccggtat gcggatcggc cccgagccga caacagactc
      961 tttcatcgcg gtaatgcacg gcgaacagga tggggtgata ccgggtaatg cccttgttgt
     1021 cgacccaaag aagccttttc gcaaactcaa cgcgtttgga aacgcatttc tcaacaggtt
     1081 catgtgtgca cagttgccta actctgtcct ggaaagcatc agtatcattg atactcctgg
     1141 aatcctgtct ggggaaaagc agaggataag cagaggttat gattttgccg ccgtgcttga
     1201 atggttcgct gagcgtgtgg accgcatcat cctcctgttc gacgcccaca aactcgacat
     1261 ttcagacgaa ttctctgagg tgatccgagc tctcaagaac cacgaggaca aaatgcgtgt
     1321 ggtgctgaac aaggcagatc agatcagcac tcagcagctg atgagggtct acggagccct
     1381 gatgtggtct ttgggaaaaa tcatcaacac acctgaggtg gtgcgtgtgt acattgggtc
     1441 gttctgggcc cagccccttt tggttcctga taacaggaag ttatttgagg cggaggagca
     1501 ggatctgttt ctggacatcc agtctctgcc acgcaacgcc gctttgcgca aactcaacga
     1561 cctcatcaaa cgagcacgcc tcgccaaggt tcaggcctac atcatcagct ctctgaaaaa
     1621 agagatgccc agtgtgtttg ggaaagacaa caagaagaag gagctgatag ccaatctggg
     1681 agagatctac accaagatcg agaaggagca ccagatctca cctggagatt ttcccaacct
     1741 tggaaagatg caggaactgc tgaatagcca ggacttcacc aaatttgcag ggctgaagcc
     1801 caaactgctg gaagcagtgg aggacatgct tgccaatgac atcgcccgtc tcatggccat
     1861 ggtgcgtcag gaggaggcga ccatgcccag tcaggctgtt aagggcggag cctttgaggg
     1921 aaccatgagt ggtccctttg gccacggcta cggagaaggt gccagcgaag gcatcgatga
     1981 gctggagtgg gtggtgggtc gcgacaagcc cacgtacgat gagatcttct acaccctctc
     2041 gcccatcaac ggtaaagtat ctggggccgc ggccaagaag gagatgctga agtccaaact
     2101 gcccaacaca gtcctgggaa agatctggaa gctagcagat gtggataagg atggtttcct
     2161 tgatgatgag gagtttgccc tggccaacca cttgatcaag gtgaagctgg agggccatga
     2221 gctgccacct gcactgcccg aacacctggt gcctccatcc aagcgtggga cggtgcagaa
     2281 cggcgacgac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2341 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2401 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2461 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2521 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2581 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2641 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2701 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2761 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2821 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2881 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2941 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3001 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3061 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3121 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3181 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3241 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3301 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3361 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3421 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3481 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3541 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3601 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3661 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3721 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3781 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3841 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3901 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3961 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4021 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4081 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4141 ttgctggcct tttgctcaca tgtt
//
