LOCUS       sequence-329890-        5331 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Gateway entry vector for N. furzeri DHTKD1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5331)
  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 5331)
  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-DHTKD1
FEATURES             Location/Qualifiers
     source          1..5331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(66..152)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     201..223
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     215..232
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     216..232
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    248..347
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3135..3234)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3251..3270)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3252..3270)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3275..3291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3275..3291)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3393..3412)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3404..4213
                     /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(3471..3490)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4360..4948
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4849..4868
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5102..5119
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5278..5305)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
       61 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
      121 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
      181 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct
      241 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
      301 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggcttcg aaggagataa
      361 caaaatgtcg gcggtgcttc ttctgaaaac gttgctgtgc aggctgccac cgagagcctc
      421 tcggcaagtc ttcggctgct tttaccacac cgagagaggc gtgtacgggt accaacccaa
      481 gcaggttgtc cgggaccaaa cgttgcagct cgagcgtatc gctgctctaa accaagatca
      541 tggtctggct cgtctggtgg aggcatacag agtacatgga cacaaggctg ctaagattaa
      601 ccccctactt ccccaccagc cggttgttga cacagtccag gagatccaca tgctgtccgg
      661 taccatgagg ggaacgcgca acacatcagg tctgcggcac ttcggacagc cccaggctcc
      721 aatagaagaa gttcaggcct acctagagaa agtttactgt gggttcctgt cggtggagac
      781 cagccagctg agcagcatgg aggagaggga gtggttcgct gaccgctttg aggagctcaa
      841 gagcaaaagc ttctctccag gtgagaggaa acacctggcc aagctcatgc tggagtctca
      901 ggaattcgat cactttctgg ccactaagtt tgcgacagtg aagcgttatg gaggagaagg
      961 agcagagagc atgatggggt ttttccacga gcttttccac cagtcggccc acagcggcgt
     1021 taccgacatc gtcattggga tgcctcacag gggccggctc aacctcctga ccggcctcct
     1081 ccagtttcca ccagagctga tgttccgtaa gatgcgtggc ctcagcgagt tccccgatac
     1141 ctcgccttcc atcggggacg ttctgtccca cctcacctcc tcggtggagt tggatgtagg
     1201 agcaggacac ccccttcatg tcaccttgct cccgaacccg tctcacctgg aggccatcaa
     1261 ccccgtggct cagggcaaaa cccgcgccag gcagcagctc aggaaagagg gcgactattc
     1321 ccccgaggaa aacgctcagc ctggggacca ggtcatctgt ttacaggtcc atggggatgg
     1381 ctccttcacc ggtcagggca ttgttccaga aaccttgact ctttcaaatc tcccccacta
     1441 cagagttggt gggagcatcc atctcattgt gaacaaccaa gtgggttata ccactccgtc
     1501 agaaagaggg aggtcctctc tgtactgcag cgatgttggt aagatggtga actgtgccgt
     1561 gatccatgta aatggagatg aagctgagga agtgctgagg gccactcggc tggccgtgga
     1621 ctaccagcga catttcagaa aggatatcat cctggatctg atctgctacc ggcagtgggg
     1681 ccacaacgag ctggatgagc ctttcttcac caacccggcc atgtacaaga tcatcaggtc
     1741 ccgtaagagc gtccctgact cctacactga ccagctgatc tcagagggtc tgatgaacga
     1801 ggaggagcgg gccgggatca agtccaaata tttcagcatg ctcaacgaaa agctgtccaa
     1861 catgacgctg tacagtcctc cgcccaccaa cctgcagggt cgctgggggg atctggtgga
     1921 gcctcaggcc agagtcacca cctgggacac gggggtcccc gtcccactgc tgcagttcat
     1981 tggtgcaaaa tcggtggaca tccctgaaca agtgcagctg cacagccacc tgctgaaaac
     2041 ccacgtacag gctcggctgc aaaagctaga ggccggatcc aaactagact ggtccacagc
     2101 agaagccctg gcatttgggt ctcttctcat tcagggcttt aatattcgaa taagtggaca
     2161 ggacgttggg agaggaacgt tcagtcagag acacgccatg gtggtttgtc aggacaccaa
     2221 cgacacgtac atccctctca accacatcag cccccagcag acggcgtttc tggaggtgtg
     2281 taacagcccg ttgtctgaag aggcggttct tgggtttgaa tacggaatga gcatcgctca
     2341 gccgaagctc ctgcctatct gggaagctca gtttggagat ttcttcaatg gagcgcagat
     2401 catctttgat acgttcatat ctggaggcga agctaagtgg ctgcttcagt gtgggatggt
     2461 gatcctgctg cctcacggct acgacggagc tggacccgaa cactcgtcct gccggatgga
     2521 gcgcttcctc cagatgtgtg acagtaaaga ggagggggtg gatggtgaca gtgtgaacat
     2581 ggcagtggtg aaccccacca cctccgcaca gtacttccac ctgctcagga ggcagatggt
     2641 tcgaaacttc cgcaaacctc tcatcgtggt gggacccaag atgctgctca gattctctgg
     2701 agcagcgtcc agcctgagtg aactagcacc aggaacagcc ttcaggccgg tgttgggaga
     2761 cgattctgtc ccagcagaaa gtgtccaaaa ggtggtgctg tgttctggga agcattacta
     2821 tgccctgctg aaacagaggg agacctcagc agccaatcag agtgcagcaa tcctccgttt
     2881 ggaggagttg tgtccgttcc ctctgcacac tcttcagcag gagctcaaaa agtaccctaa
     2941 tgccaaagag tttatttgga gtcaggaaga gcctcagaac atgggtccct ggtccttcgt
     3001 ggcccccagg tttgagaagc agctggcctg taagctccgg ttggtgagtc gaccggccct
     3061 gcccgcccct gctgtcggta tcgggaccct tcaccatcag cagcaggagg ccatcctcac
     3121 agctacgttt tccgacccag ctttcttgta caaagttggc attataagaa agcattgctt
     3181 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     3241 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     3301 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     3361 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     3421 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     3481 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     3541 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     3601 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     3661 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     3721 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     3781 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     3841 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     3901 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     3961 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     4021 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     4081 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     4141 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     4201 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     4261 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     4321 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4381 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4441 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4501 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4561 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4621 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4681 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4741 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4801 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4861 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4921 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4981 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5041 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5101 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5161 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5221 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
     5281 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta t
//
