LOCUS       sequence-413342-        4260 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 4260)
  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 4260)
  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-kcnk10a
FEATURES             Location/Qualifiers
     source          1..4260
                     /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(1869..1968)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1985..2004)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1986..2004)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2009..2025)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2009..2025)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2127..2146)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2138..2947
                     /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(2205..2224)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3094..3682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3583..3602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3836..3853
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4012..4039)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4131..4217)
                     /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 aaatttccaa cggaaaaccc
      181 gaggaagccg ggcaactgga accctccacc tgtgccagtg cagaccaacc tggtgccacc
      241 aaagaaggtc cagccaggga tgctgcagtc cagcttggtg caggccagcg tggccaccat
      301 gcagaacccc atgggctgct cattacctcg gctttctgtc tcccgaccag ctagtatggt
      361 ggccagcatg gaagcggtgg cggacggttc agcccttctc acggtcatga agtggaagac
      421 cgtgcttgcc gtgtttgtgg tggtggtagc ctacctggtt gccggcggtc tggtgttcag
      481 agctctggag caacactttg agcgctacca aaaagactct atcactttga agaaagctgc
      541 cttcctgctg aagcatccct gcgtgacccc cgatgagctg gaagagctca tcaagcattc
      601 tgtagatgca gtgaacgctg gagtcagtcc tattggagac acctcctata actccagtca
      661 ctgggactta gggagttcct ttttcttcgc tggaactgtg ataacaacaa taggttatgg
      721 taacatcgct ccaagcacag agggtgggaa aattttctgc atcctttatg caatatttgg
      781 catcccactc tttggctttc tgctggctgg tgtcggtgat caactaggga ctatttttgg
      841 gaagagcatc gctaaagtgg aaaaaatgtt caggcgcaaa cacaatcaaa taagccaaac
      901 caagattcga gtggcctcta cactgctctt catcctggcc ggctgcatcc tcttcgtcac
      961 cataccagcc attatcttca agcacattga aggatggact gggctagagg ccatttattt
     1021 cgtggtcatc accctcacga ctgttggaat tggagattat gtcgcaggag gcaatagacg
     1081 aatagagtat cgcaagtggt acaggccact ggtgtggttc tggatcctgg ttggactggc
     1141 ttactttgca gcagttttaa gcatgattgg agactggctg agggtgctct ctaaaaagac
     1201 aaaaatggag gtgggtgaaa ttaaggctca cgctgctgag tggaaggcca atgttcgagc
     1261 ggagctgcgt gagaccaggc gacgactcag tgtagaagtt cacgataaac ttcagagggc
     1321 tgccaccatc cgcagcatgg agcgcaggca gctgggattt gaccagcgtg cccattccct
     1381 cgacatgttg tcccctgaac gacgagctgc cttcaacagc ctggatgcca ccaacttcaa
     1441 gacatcctcc caggagagta tcgacacaaa gctcaacaac ctgcgtctga gagtagaaca
     1501 gaacgagcat cgcaggagcg accctagcca ggcgtactct gaagacaaca tttttaaccg
     1561 gcttggttca gtcaccaaac tcgccaagcg gaacaagaac agggatttga agaaaaacat
     1621 cctggatgat ggtcggaaag ccttagattc cttcagctgt gacactcccc ctatggacga
     1681 agagaagaag gaggccgagg atgaggagct tgagaaagag gtcaacatta gtctcaccaa
     1741 cttgcctctc tttgtggaga gtcccaacaa acaaaatgga ttcgtgcctc ttccgccaca
     1801 gaccaaagag gaagaaacag agacaaaact tgaggataaa gagttccgtt tgcaggtgga
     1861 tccataggac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1921 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     1981 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2041 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2101 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2161 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2221 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2281 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2341 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2401 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2461 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2521 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2581 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2641 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2701 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2761 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2821 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2881 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2941 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3001 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3061 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3121 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3181 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3241 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3301 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3361 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3421 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3481 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3541 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3601 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3661 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3721 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3781 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3841 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3901 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3961 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
     4021 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
     4081 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
     4141 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
     4201 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
//
