LOCUS       sequence-330247-        4716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for N. furzeri MARK4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4716)
  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 4716)
  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-MARK4(v3)
FEATURES             Location/Qualifiers
     source          1..4716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..43)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(26..42)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(35..57)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      106..192
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      284..311
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(470..487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(641..1229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(721..740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1376..2185)
                     /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     2099..2118
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2177..2196
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2298..2314
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2298..2314
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2319..2338
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2319..2337
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2355..2454
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(join(4627..4716,1..10))
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tcattatttg gggcccgagc ttaagactgg ccgtcgtttt acaacgtcgt gactgggaaa
       61 acatccatgc tagcgttaac gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
      121 gctcagtcgg aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
      181 agtaggacaa atccgccggg agcggatttg aacgttgtga agcaacggcc cggagggtgg
      241 cgggcaggac gcccgccata aactgccagg catcaaacta agcagaaggc catcctgacg
      301 gatggccttt ttgcgtttct acaaactctt cctggctagc ggtacgcgta ttaattgcgt
      361 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
      421 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
      481 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
      541 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
      601 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      661 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      721 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      781 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
      841 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
      901 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
      961 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1021 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1081 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1141 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1201 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1261 acgctcagtg gaacgacgcg taactcacgt taagggattt tggtcatgag cttgcgccgt
     1321 cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga
     1381 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     1441 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     1501 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     1561 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     1621 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     1681 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     1741 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     1801 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     1861 ctaatacctg gaatgctgtt tttccgggga tcgcagtggt gagtaaccat gcatcatcag
     1921 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     1981 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     2041 ctggcgcatc gggcttccca tacaagcgat agattgtcgc acctgattgc ccgacattat
     2101 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg
     2161 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     2221 gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     2281 acacgggcca gagctgccag gaaacagcta tgaccatgta atacgactca ctatagggga
     2341 tatcagctgg atggcaaata atgattttat tttgactgat agtgacctgt tcgttgcaac
     2401 aaattgataa gcaatgcttt cttataatgc caactttgta caagaaagct gggtcaagtt
     2461 caaggtcttt agagatctgg gtggcaatgt cccgaaaagc tagcggtgcc ccccagagtc
     2521 gcgtgtagcg caccccgtta gaagtgtcgg aggtgcccgt ggagagatgg cacacctcgg
     2581 cctcaaaagc cacacggccc cctgccatgc catgggtgca agagagcaga aaggggccag
     2641 aatggtggac ctggcagcca catccgagtg ctgcctcccg cagagccaga acaacctccg
     2701 cagggtcccg ccgcccaccg cctctcagat tccagccacg tccgatggcc cggggctcgg
     2761 acgctttttg atacccaggt agatggcaac ttgtgaccgc aagtctcctg attccctcag
     2821 attcatttgt gacctaactg tttttgatcc ttgtggaaga gtacaacccg agactgattt
     2881 gtttgagcct tggcgcttgg acgggtctag gttgaccctg cgagtgagtt ttgaggtcag
     2941 cttggtgaag aggttggagg ttgccctggt gcgagcatgg ggcaacgggc tggcttcatg
     3001 gtgggacagg gtgggtgatg tgggtggggc agagtagacg ggaggaccac ggtctctcag
     3061 ctgtccccca tggaaggtgc tgcggatggt ggaccctcgg gtcaaacggc accggtccga
     3121 agaggacgag gaggaagctg ctccagctcc agagatgctg agcgtggagg gagaagctgc
     3181 aggcaggcgg tgacccagag agctgttttc tttgccattc tgcaggagtg agtgtctgtc
     3241 agcgcccgag cggtccgtgc acacgtaggt gttccggcgt gtcatggtgc tcccagggat
     3301 gttattagaa gtcgctgtgc cgtcgctgcc tgtggtaggt ggaggcactg cgctgggtct
     3361 tgctatgcga agaggaggtg gaggaggagg aggagccagt ggctgaggag tgtttgttgg
     3421 tcccgttagt gatcgggctg ggtcgtaccc tcgccaagga caggctacta gcagagcggg
     3481 catcactgct ctcatcgcct tttcggccca ggagcaaata tgtggcggtg acctcgttgt
     3541 atttctgatt gagcagagag tctttgattt cctcgggggt gaagcccatc ccgaccatca
     3601 cctctatgcg agccggatca ctgtagtcct ccacaggctc tatgtggggc ttcaggtcgt
     3661 caccctcgta ccctgtgttt atccatttgt ccttcatgac ttgctccagc gtgcagcgct
     3721 tggctggatt gagcaccagg aatcggcgaa ggattccctc acagtcagtt gacatgtaga
     3781 agggcactcg gtactttcct ctcagcacac gctccctcag ctccttgagg ttctgtccat
     3841 caaaaggcag cgagccgctg accagagtgt agaggatgac tcccagactc cagacatcca
     3901 cctcgggccc atcatacttc ttcccctgga agagctcggg ggcagcgtag ggcggcgagc
     3961 cacagaacgt gtccagcttg tttcccagag tgaactcatt gctgaagcca aaatcagcta
     4021 tcttgatgtt agcgtcggca tccagcagaa ggttctccgc cttcagatct cggtggacta
     4081 tgttcttggt gtggcagtag tggacagcag agacgatctg acgaaatttg gctctggctt
     4141 caacctcctt cattctccca tgagacacga ggtagtcaaa cacttcccct ccactggcat
     4201 actccatgac caggtaaagg gtcttatccg tctcaatcac ctcaaacagc ttcactatgt
     4261 tagggtgatt taggcccttc attattcgca cctcacggaa aagcttttgt aggctggttg
     4321 ggttcaactg agttttatcg atgattttta ttgcaacctc tctgcctgtc agaatgtggc
     4381 gagccagttt gactttggca aagtttccct tgccaatggt cttgagcaga cggtagttgc
     4441 caatgtgtgg ctgctcatca gagcaggagg cgatggagtt cctgcagcgg gcgcccagag
     4501 agcggctgga ctggctcgtg gccttgtctg aacggctggc ccccagcgac acatgatgtt
     4561 cccctgtgct cctgtcgctc ccagacggta acgcagatcg agaggacatt ttgttatctc
     4621 cttcgaagcc tgcttttttg tacaaagttg gcattataaa aaagcattgc tcatcaattt
     4681 gttgcaacga acaggtcact atcagtcaaa ataaaa
//
