LOCUS       sequence-413259-        3624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Riboprobe generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3624)
  AUTHORS   Park SJ, Silic MR, Staab PL, Chen J, Zackschewski EL, Zhang G
  TITLE     Evolution of two-pore domain potassium channels and their gene 
            expression in zebrafish embryos.
  JOURNAL   Dev Dyn. 2024 Jan 25. doi: 10.1002/dvdy.690.
  PUBMED    38270285
REFERENCE   2  (bases 1 to 3624)
  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"; journalName: "Dev Dyn. 
            2024 Jan 25. doi: 10.1002/dvdy.690."
COMMENT     Sequence Label: pDONR221-kcnk15
FEATURES             Location/Qualifiers
     source          1..3624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..28
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     20..37
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     21..37
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    53..152
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1233..1332)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1349..1368)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1350..1368)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1373..1389)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1373..1389)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1491..1510)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1502..2311
                     /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(1569..1588)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2458..3046
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2947..2966
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3200..3217
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3376..3403)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3495..3581)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat
       61 gattttattt tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt
      121 tataatgcca actttgtaca aaaaagcagg cttcaccatg aagaagcaga atgtgcgcac
      181 tctgtccctc atcctctgca tcttctccta tctgctggtg ggcgccgcgg tgttcgtggc
      241 gctggagtcc gacaccgaga gcgcccgaaa gcggatgctg gagcacaagc gcgcggagct
      301 gcggaggaaa taccgcttta ccgacggaga ctaccaggag ctggagcgcg tgctgcggca
      361 ggcggagcca caccgcgccg gcacacagtg gaggttcgcc ggatcctttt acttcgccat
      421 caccgtcatc accaccatag ggtacgggca cgcggcccca ggcacagacg cagggaagct
      481 gttctgcatg ctgtacgccg gtctgggcat cccgctgacc ctggtgatgt tccagagtct
      541 gggcgagcgc atgaacaccg gcgtgcggtt cctcctcagc cgcatgaaga gagctctggg
      601 cctgcagcgc accgagatct ccacgcagaa catggtgctg gtgggtgtgc tgtcctgcct
      661 gggcacgctc tgtgtgggcg ccgctgcatt ctcacacttc gagagctgga cgttcttcca
      721 tgcctactac tactgcttca tcacgctcac caccatcggc ttcggggact tcgtggctct
      781 ccagaagaag gaggacctgc aggagaacca gccgtacgtg ctgttcagct tcatctacat
      841 cctgctgggg ctgacggtga tcggggcctt cctcaatctg gtggttctgc gcttcctgac
      901 gctaaacact gaggacgaga agcgggacgc agcggagcgg gcatcgcagc gggcatcact
      961 gcgggcatcc tcactgcgca aggctccagc tttcagacgg gacagccaga cacacagccg
     1021 cacaaacctg ctgtccccgc ggcccacacc gcacctgcgc tggttctgca gccgcgtaca
     1081 cctggatcac acctccgcat gcagaaacac ctgtgccggc tgcagcgtga gctccgtgtc
     1141 ctgcaggatt catgcggaca ctgcgttctc ctccccgcgc agcacactat ctgctggact
     1201 cagcttcacc acacgcagga aatctctgta agacccagct ttcttgtaca aagttggcat
     1261 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     1321 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     1381 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     1441 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     1501 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     1561 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1621 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1681 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     1741 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     1801 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     1861 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     1921 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     1981 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2041 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2101 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2161 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2221 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2281 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     2341 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     2401 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2461 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2521 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2581 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2641 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2701 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2761 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2821 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2881 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2941 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3001 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3061 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3121 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3181 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3241 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3301 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga
     3361 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg
     3421 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa
     3481 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     3541 cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac
     3601 tctcgcgtta acgctagcat ggat
//
