LOCUS       sequence-413260-        3471 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 3471)
  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 3471)
  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-kcnk20
FEATURES             Location/Qualifiers
     source          1..3471
                     /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(1080..1179)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1196..1215)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1197..1215)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1220..1236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1220..1236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1338..1357)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1349..2158
                     /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(1416..1435)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2305..2893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2794..2813
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3047..3064
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3223..3250)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3342..3428)
                     /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 gcagcaccag ctacccacaa
      181 tggaaaaatc ttcctaatct tctatgggct gataggctgc gcaaccacca tgctattctt
      241 caacctcttt ctggagcgca tgataacact gattacattc ctcacgaaca gatgtcgaaa
      301 gcaacaacaa aggactaaac agagtaaaag acacattgtc ccaaatgtaa acacacatcc
      361 agaaaatgga gacatccacg agtcatggaa accatccgtt tactatgtta ccctcattct
      421 ggcagtagtg gcactgctgg tgaacttggg ggcatcaggc ttgtattcag ccatggagga
      481 ttggagctat ctggagtcga tgtacttctg ctttgtggct tttaccacca tgggtttcgg
      541 ggacatggtg agcggacaga aggcctatta tgaagtacgt tgggtttacc aggttgccaa
      601 ctctttaatg atcatccttg gcgtcagctg cacctactct cttgtcagcg tcaccgctat
      661 catcatcaaa cagatgttga actggatttt ggccaagctg ttcagcatgc actgctgttt
      721 ctccatcact actccaaaag tcaaagaagc agccaatcag cagatccaga acgcgagcag
      781 ccatcagagc acctgccatg tggtcaaggc ctcttgctcc tccaagagca agtgtaagtg
      841 cgccagctct gcagtagaaa ctgtgtgcca gagcggggac acagcaaaca gcgctccaag
      901 ctttgaacgc caaatcagac attcggacag actgtcagag gatggtgcag caaaagagct
      961 ctgcagtgct gtcagcaaga accgtcatct ctgtgttatg caggaggatg tgagaacagt
     1021 tgcactgtca aatcagcagc agcaagagac aaacattaat cctagtccca tactttaaga
     1081 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     1141 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     1201 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     1261 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     1321 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     1381 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     1441 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     1501 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     1561 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     1621 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     1681 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     1741 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     1801 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     1861 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     1921 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     1981 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     2041 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     2101 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     2161 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     2221 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     2281 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2341 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2401 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2461 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2521 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2581 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2641 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2701 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2761 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2821 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2881 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2941 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3001 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3061 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3121 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3181 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     3241 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
     3301 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
     3361 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt
     3421 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga t
//
