LOCUS       sequence-413330-        3993 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 3993)
  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 3993)
  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-kcnk19/kcnk12l
FEATURES             Location/Qualifiers
     source          1..3993
                     /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(1602..1701)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1718..1737)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1719..1737)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1742..1758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1742..1758)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1860..1879)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1871..2680
                     /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(1938..1957)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2827..3415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3316..3335
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3569..3586
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3745..3772)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3864..3950)
                     /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 ccatgaaagt
      181 ggctgtgcat gtttgcctgt tcttgtcatt tcctgctgat ggtcaagtca tgccacgtcg
      241 aagagatgcc agcaacaact gttgcctccc acttcacctg aatgaagaca atgcccgttt
      301 cggcttgttg gcagccctca tcctgctcta cctgctgtgt ggcgcggtgg tgttctccgc
      361 acttgagcat ccctctgagc tgctggctca ccagcgctgg gaagaacagc tggccaactt
      421 cacagagcaa aacagtgttc atctcaaatc actgcaggtc ctgctcaggc aatatgagga
      481 ggcctttgca gctggaatcc gggtggacaa acttagggcc cgctgggatt tctcaggcgc
      541 cttctacttt gttggtacag tgatttccac tataggtttt ggcatgacca ctcctgtcac
      601 cgttgcaggt aagatctttc tgatattcta cggactcctc ggctgtgcag caacaatcct
      661 cttcttcaac ctgtttctgg agcgcatcat cacgatgctg gcctacatta tgcgctggtg
      721 tcatgagcgc cagcttcggc ggtccggcgt gggaggagag gaggccagaa gtgaggacga
      781 cagtctggag ggctggaaac cttcagtcta ctacgtgatg ctcattctgg gcatcgctgc
      841 cctgctgatc gcatgcagcg cgtctgcttt gtactctgcc atggagggct gggactactt
      901 tgaatccttt tacttctgct ttgtggcctt cagcactatt ggatttgggg atgtggtgag
      961 cagccagcgg gaaaactata aagctcagga ggcctaccgt ttcggcaact gcctcttcat
     1021 cctgatgggt gtgtgttgca tttattccct gttcaatgtc atttccatca ttatcaagca
     1081 gacgctcaat tggatcctcg gcaagctgga ttgcagtagg acccagtgtc cctgtcgtgg
     1141 tcgtccatac aggcggcgag gtcgggcctg cggttgctgc tgttttccac gtattcccaa
     1201 ccagcgccac aaccaccatc ctccaccagg aaattcccga cagagggcac aaaaacgtaa
     1261 cgctgtgcac cctgccccag caaatgaagc gtcaggaaag cgcttcacta atgcatcagt
     1321 ggaaacagtt tgtgatagtg agactgatgc tggcttagga ccagatggag gttatctgac
     1381 agggcgcaga ctgtctggag aaatgatctc agtcaatgac tttatggcca acaaggtttc
     1441 tctggccatt ctgcagaagc aactctcaga gaccgctcac ggtaatccaa gacagagcca
     1501 tgttcgccag aatggctttt ctggtggagt gggagctttt gctatcatga ataaccgcct
     1561 tcaggagacc agcgtagaca ggtagaaggg tgggcgcgcc gacccagctt tcttgtacaa
     1621 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1681 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     1741 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     1801 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     1861 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     1921 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1981 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2041 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2101 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2161 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2221 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2281 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2341 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2401 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     2461 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2521 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2581 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2641 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2701 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     2761 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2821 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2881 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2941 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3001 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3061 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3121 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3181 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3241 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3301 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3361 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3421 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3481 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3541 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3601 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     3661 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag
     3721 ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt
     3781 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca
     3841 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
     3901 cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca
     3961 gttccctact ctcgcgttaa cgctagcatg gat
//
