LOCUS       sequence-413284-        3723 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 3723)
  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 3723)
  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-kcnk3a
FEATURES             Location/Qualifiers
     source          1..3723
                     /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(1332..1431)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1448..1467)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1449..1467)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1472..1488)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1472..1488)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1590..1609)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1601..2410
                     /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(1668..1687)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2557..3145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3046..3065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3299..3316
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3475..3502)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3594..3680)
                     /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 aagagacaaa acatcaggac
      181 cctcgtgttg atcatctgta cttttactta tttgctcgtc ggagcggcgg tttttgatgc
      241 attagagtct aagatggaga tcacccagaa gaagatcctg gacgatcgga agcgagagct
      301 gatggacaag tacaacttga gcaaagtcaa tttcgacgag ctggagcggg tcgtgctgca
      361 gctcaagccg cacaaggcgg gggtccaatg gaggttcgct gggtccttct atttcgccat
      421 cacggtgatt acgactatag gatatggtca tgcggccccc agcacggatg gcgggaaggt
      481 gttctgcatg ttctatgctc tcctggggat cccgctcacc ctggtcatgt tccaaagtct
      541 gggtgaacgc atcaacacct tcgtcaaata cctgcttcac cgtctcaaaa agtgtcttgg
      601 tctccgtcac acagaggtct ccatggccaa catggtgtgc attggcttga tatcctgcat
      661 gagcaccctg tgtgtggggg ctgcggcttt ctcccgctac gaggactgga gcttcttcca
      721 tgcatactat tactgcttca ttacactcac caccattggt tttggggact acgtggccct
      781 gcagaaggat aacgctttac aaaccaaccc ccagtatgta gcctttagct tcatgtacat
      841 cctcacaggc ctcactgtca ttggagcctt cttgaacctg gtggttctgc gttttatgac
      901 catgaatgca gaagatgaac gccgtgacgc agaacagcgc actttgctgt cccgaaatag
      961 gcagagcgga ggcgccggaa ccaatcattg catttccgac ccaccttctt catctaccgc
     1021 gggacacgga ggagggtccg gtaaggggtt gtgcaacgtc tacgcagaag tgctccattt
     1081 ccagtccatg tgctcttgtc tttggtacaa aagccaagag aagatgcgct actcaatccc
     1141 catgattatc tctcacgatc tgtccacttc agacacctac atggagcaca gcgagacctc
     1201 agatgtcctt cactccaatg gctgtgtgtg cagtgccttg caggagcact ctgccgccag
     1261 ttcagtgtac acaggcctgc ttagcccagc acattaccaa agactctcca aacgtcgcag
     1321 ctctatttag gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     1381 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     1441 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     1501 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     1561 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     1621 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     1681 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     1741 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     1801 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     1861 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     1921 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     1981 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2041 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2101 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     2161 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     2221 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     2281 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     2341 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     2401 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     2461 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     2521 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2581 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2641 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2701 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2761 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2821 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2881 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2941 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3001 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3061 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3121 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3181 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3241 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3301 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     3361 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     3421 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
     3481 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
     3541 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
     3601 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
     3661 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
     3721 gat
//
