LOCUS       sequence-413329-        4038 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 4038)
  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 4038)
  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: pENTR-D-Topo-kcnk4a
FEATURES             Location/Qualifiers
     source          1..4038
                     /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(1647..1746)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1763..1782)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1764..1782)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1787..1803)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1787..1803)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1905..1924)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1916..2725
                     /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(1983..2002)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2872..3460
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3361..3380
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3614..3631
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3790..3817)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3909..3995)
                     /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 ctccgcggcc gcccccttca ccatgtccaa
      181 ggtgttgcag aaagtggttg acgccattga ggcaggtgtg gatgccagaa gcacatcaaa
      241 ttacaccagc agatgggatc catccaatgc tttctttttc tgtggcacca taatcaccac
      301 catcggcttt ggaaacatct cacccaagac caaaggtggt cagttgttct gcatatttta
      361 cgctctggtg gggatcccca tgtttgggat actgttagcc ggggttggag atcacctggg
      421 caccctgttg cggagagccg tggccaaggt ggagactttg ttcttgcgca agggtgtgaa
      481 gcccaccagt gtccgcataa tctcggctgt gttctccatc ctgattggct gcttggtctt
      541 catcgccgtg ccgaccatgg tgtttcagga agtggagagc tggagcctgc tggaagcggt
      601 gtattttgtt gtgatcacac tcaccacggt gggcttcggg gactatgtgg cagagaacag
      661 gagagatgga acgcccctgt acaagccatt ggtgtggttg tggatagtgt ttggtctggc
      721 gtattttgca tccatcctca cgatgattgg gaactggctt agagtccttt caaagaagac
      781 gcgtgcagag atggaggagc tgagagctca cgccacagac tggactcaaa acatccagaa
      841 catgtctatg gacttccgta ttcccgttcc actggatctt aatgacccct ttcagcttca
      901 gcggagacgc agaaggaagc gccaccacct ccaacatcat catcatcgcc acccaagacc
      961 aaacacaaca ggactgactc atgggatctc tcttgatgcc gatattatcc gagagaacgg
     1021 acatctgatc cttggctggc cgacatgcag atacaaacca gcttcagaag tactgtccca
     1081 atccaggtca atgagcagga ttgagggagc gcattatgag atgagaccag gatcacggct
     1141 gacagtagga ccgacgtcga gatccatctc acggctagag ctaaaaccag catccagagc
     1201 cgggtccaga gccatttcca gatccccctc acaatccgaa tcggggtcat tttctgcatc
     1261 cgagacacca tctgagtcct ggtcggaata tgaatacgag tcaaactcct gtaactcaga
     1321 agatgtggaa tctggagcga aggcatcacg gagatcactg gaaagtcaga atgatgttcc
     1381 gattgtggta gttgcaagct gcagtccacc ccgtccgtcc cttctggact tctttggcga
     1441 gaacctagcc tacatcgacg agtcttccga cgctcttagt gatcgggtca aaccgtcagg
     1501 acagaaccgt caacgcagac caaagaggag gagtatgaca aagcggcagc ctgctggact
     1561 acagagggag gtcagtaatg aactacaacc cccatcacat cctccgactc caccacctcc
     1621 aacatcttag aagggtgggc gcgccgaccc agctttcttg tacaaagttg gcattataag
     1681 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     1741 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     1801 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1861 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     1921 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     1981 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2041 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2101 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2161 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2221 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2281 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2341 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2401 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2461 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2521 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2581 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2641 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2701 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     2761 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     2821 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2881 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2941 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3001 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3061 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3121 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3181 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3241 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3301 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3361 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3421 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3481 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3541 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3601 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     3661 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     3721 gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt
     3781 agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt
     3841 tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct
     3901 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
     3961 gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
     4021 gttaacgcta gcatggat
//
