LOCUS       sequence-413266-        3702 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 3702)
  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 3702)
  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-kcnk3b
FEATURES             Location/Qualifiers
     source          1..3702
                     /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(1311..1410)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1427..1446)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1428..1446)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1451..1467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1451..1467)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1569..1588)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1580..2389
                     /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(1647..1666)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2536..3124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3025..3044
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3278..3295
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3454..3481)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3573..3659)
                     /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 aagaggcaaa acgtgcggac
      181 actcgcctta atcatctgca ctttgtccta tttgctcatc ggagcgggag ttttcgacgc
      241 tctcgagtca aagcaagaga aaagccagaa gggcagactc gactaccgca aattcctgct
      301 catgcataaa tacaacctca ccaggcttga ctttgatcag attgagaagg tcgtgttgct
      361 tctgaagccc cacaaagcag gagtccagtg gaagttctcc gggtctttct atttcgccat
      421 cactgtgatt acaaccatag gttatggcca tgccgccccc agtacggatg ccggtaaggc
      481 attctgcatg gggtatgcac ttctgggcat ccccctcacc ctggtgatgt tccagagcct
      541 gggcgaacgc attaacacct tcgtccgctt cctgttgcac aaagccaaga aatgcatggg
      601 cctgcggcgg ccggaagtct ccatggccaa catggtgatc atcggctttt tctcctgcgt
      661 cagcaccttg tgcataggag cagctgcctt ttcccactac gaaggttgga cctttttcca
      721 tgccttctac tactgcttca tcaccctcac caccatcggt tttggcgact acgtagccct
      781 acaaaaggac aacgctctcc agaacgaccc tcattacgtg gccttcagtt ttgtctacat
      841 tctgatgggc ctcacagtca tcggtgcttt cctcaattta gtggtgctac gcttcatgac
      901 catgaacaca gaggacgaga gaagagatgc cgagcagaga gcgctgctct ccaaagataa
      961 acaccaaggt ccagtattac gccgtgcaga acctccgagc ccatccgctg tgggccgaga
     1021 caaaagacga gggctgaaga gcgtctacgc cgaggtgctc cacttccaga cggtgtgctc
     1081 ctgcctgtgg tacagaagcc gagagaaatt ggtcatcctc ccacaggatc tgtcattcac
     1141 tgatgcgctt atggagcaag gagatgtatc gccccatcac ttcttcgaaa acggccctac
     1201 gggctgcgtg tgtaaccctc aacgctgctc ggccattagc actgtgtctg ctgacctgag
     1261 aaacatctcg ccgttcaggc tcttctccaa aagacgcagc tctgtctgag acccagcttt
     1321 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     1381 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     1441 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     1501 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1561 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     1621 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1681 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1741 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1801 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1861 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     1921 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1981 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2041 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2101 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     2161 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2221 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2281 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2341 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     2401 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     2461 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     2521 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2581 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2641 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2701 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2761 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2821 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2881 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2941 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3001 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3061 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3121 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3181 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3241 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3301 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3361 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
     3421 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     3481 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     3541 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     3601 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
     3661 tgcctggcag ttccctactc tcgcgttaac gctagcatgg at
//
