LOCUS       sequence-329532-        3864 bp    DNA     circular SYN 15-APR-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 3864)
  AUTHORS   Yiwen R. Chen , Itamar Harel, Param Priya Singh, Inbal Ziv, Eitan 
            Moses, Uri Goshtchevsky, Ben E. Machado, Anne Brunet, Daniel F. 
            Jarosz
  TITLE     Tissue-specific landscape of protein aggregation and quality control
            in an aging vertebrate
  JOURNAL   bioRxiv
REFERENCE   2  (bases 1 to 3864)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv"
COMMENT     Sequence Label: pDONR221-MARK4(v2)
FEATURES             Location/Qualifiers
     source          1..3864
                     /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(1989..2088)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2105..2124)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2106..2124)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2129..2145)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2129..2145)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2247..2266)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2258..3067
                     /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(2325..2344)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3214..3802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3703..3722
                     /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 acaaaatgtc ctctcgatct gcgttaccgt ctgggagcga
      721 caggagcaca ggggaacatc atgtgtcgct gggggccagc cgttcagaca aggccacgag
      781 ccagtccagc cgctctctgg gcgcccgctg caggaactcc atcgcctcct gctctgatga
      841 gcagccacac attggcaact accgtctgct caagaccatt ggcaagggaa actttgccaa
      901 agtcaaactg gctcgccaca ttctgacagg cagagaggtt gcaataaaaa tcatcgataa
      961 aactcagttg aacccaacca gcctacaaaa gcttttccgt gaggtgcgaa taatgaaggg
     1021 cctaaatcac cctaacatag tgaagctgtt tgaggtgatt gagacggata agacccttta
     1081 cctggtcatg gagtatgcca gtggagggga agtgtttgac tacctcgtgt ctcatgggag
     1141 aatgaaggag gttgaagcca gagccaaatt tcgtcagatc gtctctgctg tccactactg
     1201 ccacaccaag aacatagtcc accgagatct gaaggcggag aaccttctgc tggatgccga
     1261 cgctaacatc aagatagctg attttggctt cagcaatgag ttcactctgg gaaacaagct
     1321 ggacacgttc tgtggctcgc cgccctacgc tgcccccgag ctcttccagg ggaagaagta
     1381 tgatgggccc gaggtggatg tctggagtct gggagtcatc ctctacactc tggtcagcgg
     1441 ctcgctgcct tttgatggac agaacctcaa ggagctgagg gagcgtgtgc tgagaggaaa
     1501 gtaccgagtg cccttctaca tgtcaactga ctgtgaggga atccttcgcc gattcctggt
     1561 gctcaatcca gccaagcgct gcacgctgga gcaagtcatg aaggacaaat ggataaacac
     1621 agggtacgag ggtgacgacc tgaagcccca catagagcct gtggaggact acagtgatcc
     1681 ggctcgcata gaggtgatgg tcgggatggg cttcaccccc gaggaaatca aagactctct
     1741 gctcaatcag aaatacaacg aggtcaccgc cacatatttg ctcctgggcc gaaaaggcga
     1801 tgagagcagt gatgcccgct ctgctagtag cctgtccttg gcgagggtac gacccagccc
     1861 gatcactaac gggaccaaca aacactcctc agccactggc tcctcctcct cctccacctc
     1921 ctcttcgcat agcaagaccc agcgcagtgc ctccacctac cacaggcagc gacggcacag
     1981 cgacttcgac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2041 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2101 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2161 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2221 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2281 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2341 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2401 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2461 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2521 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2581 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2641 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2701 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2761 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2821 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2881 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2941 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3001 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3061 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3121 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3181 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3241 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3301 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3361 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3421 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3481 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3541 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3601 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3661 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3721 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3781 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3841 ttgctggcct tttgctcaca tgtt
//
