LOCUS       sequence-413325-        3813 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 3813)
  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 3813)
  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-kcnk12
FEATURES             Location/Qualifiers
     source          1..3813
                     /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(1422..1521)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1538..1557)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1539..1557)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1562..1578)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1562..1578)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1680..1699)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1691..2500
                     /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(1758..1777)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2647..3235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3136..3155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3389..3406
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3565..3592)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3684..3770)
                     /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 ccatgatgcc
      181 ggcgcagaga ctgcagggct gccgctccct tcacatcaac gaagacaatg gacgctttct
      241 gctgctcgcc ctgctcatca tcgtgtacct gctgtgcggc gctgcggtgt tttccgccat
      301 cgagaggccg tccgagctgc gggcgcacgg gcgctggaac cggaccctcc tcaactttag
      361 cgacgcgttc aatatcagtt tgcaggatct gagctccttc ctgaaagagt acgaggctgc
      421 gattgctgct ggagttcggg tcgatgctct tcgacccaga tgggatttta ctggggcgtt
      481 ttactttgtt ggaacagtgg tgtccactat tggcttcggc atgagcaccc ctgtgacagt
      541 cgccgggaag attttcctga tcttctatgg tctgttaggc tgcgcggcca ccatcctctt
      601 cttcaacctc ttcctggagc gcatcattac cctgctggcg gtggtgatga aggctgtgcg
      661 tctgcgccgt ctgcgcacca gcggcctcct cccgccggga atctgccagg actttgccgc
      721 cagcacgtta ccaggctgga agccctcggt gtaccatgtc atgctgatcc tgggtctgtc
      781 agccatcgtg atctcctgct gcgcatcagc catgtatacc ccagtggagg gctgggcgta
      841 tgtggactcg ctttatttct gcttcgtcac tttcagcacc attggtttcg gggatttggt
      901 gagcagtcaa aacgctgcgt acgactacca aggactctat cggctagcca actgtctgtt
      961 tatattggca ggagtgtgtt gcatctattc tctcttcaac gtcatctcta ttgtgattaa
     1021 gcaagtcctc aattggatgt tggggaaaat gagttgcctg tgctgccaga gatgcaacaa
     1081 agctaatgct ttcttaggac gccgtaatgc tatcagaccc ggttctagac tcagacgcgg
     1141 acgattcgga cagacgccgg attccgatgg gccttgcgat agcgataccg aaggccgacg
     1201 gctttcaggg gagatgatct ccatgcggga tctgacggga tctaataaaa tatctctcgc
     1261 aattatgcag aaacagctgt cggagtctgc taatggatat cccagaacgg tgtgcggaag
     1321 ttcacgacag aatggatttt caggtggcgt cggtgcgctt ggtatcatga ataaccgatt
     1381 agcagaaacc agtgactcta ggtagaaggg tgggcgcgcc gacccagctt tcttgtacaa
     1441 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1501 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     1561 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     1621 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     1681 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     1741 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1801 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     1861 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     1921 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     1981 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2041 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2101 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2161 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2221 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     2281 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2341 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2401 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2461 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2521 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     2581 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2641 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2701 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2761 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2821 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2881 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2941 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3001 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3061 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3121 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3181 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3241 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3301 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3361 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3421 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     3481 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag
     3541 ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt
     3601 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca
     3661 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
     3721 cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca
     3781 gttccctact ctcgcgttaa cgctagcatg gat
//
