LOCUS       sequence-413328-        4203 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 4203)
  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 4203)
  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-kcnk10b
FEATURES             Location/Qualifiers
     source          1..4203
                     /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(1812..1911)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1928..1947)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1929..1947)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1952..1968)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1952..1968)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2070..2089)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2081..2890
                     /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(2148..2167)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3037..3625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3526..3545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3779..3796
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3955..3982)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4074..4160)
                     /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 gtcccgcaga agaagccgca
      181 gtcgggtgca gtccagtcca gtctggtcca ggccagtgtt tcggccatgc agaacccgat
      241 gggctgcgag ctgaaaaacg gtcactgtct actaccacga ctgtcaatct catcccgatc
      301 ggccagcgtc ctcaccggca tggattctgg agcggacgga tcggctctgc actccgtcat
      361 gaagtggaaa acggtgctgg ctgtgtttgt ggtggtgcta tcgtatttag tgtgcggagg
      421 actagcattt caggcactcg aacagccctt cgagagcatc cagaaagact ccatcaccca
      481 gaagaaagcc cagtttctcc agaaaaaccc ctgcgtctct cctgctgacc tcgaggcgct
      541 tattaagcat gcagtagaag cggtcagcgc tggagtgagt cctattggag acgcgtccta
      601 taactccagc cagtgggatc tcggcagcgc tttcttcttc gctggaaccg tcatcactac
      661 aatcggctat ggaaacatcg ctcccagcac tgagggcggc aagattttct gcatccttta
      721 cgccatattt ggcatccctc tgttcggctt cctgttggcc ggggtcgggg atcaactggg
      781 cacgatgttt atgaagagca tcctgagggt ggagaaagtt ttcaggcaaa agcacaagca
      841 aatcagtcag accaaaatcc gcgtgacctc caccatcctc ttcatcatcg ccggatgcat
      901 cgtttttgtc accatcccag cggtgttttt caagcacaca gagggatgga gtacattaga
      961 cgccatttac tttgtcgtta ttacgctcac gaccatcggc attggagatt atgtggcagg
     1021 aggagatcgg aaaattgaat acatgaaatg gtacaagcct ctggtgtggt tctggatttt
     1081 ggtgggtctg gcgtactttg cagcagttct tagcatgatt ggagattggc tcagggttct
     1141 gtccaagaaa acaaaagagg aggttggaca acaagttgga ggcattaagg ctcatgcggt
     1201 ggaatggaaa gcaaacgtcc gagcagagtt acgtgaaaca agacgccgtc tcagtggaga
     1261 aattcacgat aaactccagc gagctgcgac tattcgcagc atggagcgcc ggcgtttggg
     1321 tctggagcag cgtgcgcatt cgctcgacat gctatcgcct gaaaaacgag cggtttttgc
     1381 tagtttggat gccggacgct tcaaaacatc ctcgcaggag agcatcgaca caaaactcaa
     1441 caacctgcgc ctgaaggcta aatgcggaga gcagcagacg tcttcagaag aaaacatctt
     1501 caaccgcttc ggctcgctca ctaaactagc caaacgcaac aggagtaaag acctgcgcag
     1561 gaatattcag gaggatccaa ctcagataga cgaactcgac aacgtcacgc tgggagaaga
     1621 gagacaggag gaggaggacg aggaccagat ggaggaggag gaggaaaagg tggaaaaaca
     1681 ggccaacact ggatttacct gtctgccgca ctatcctgag gaacccaaac agcagaacgg
     1741 tttcgcaccc gctcaggtta aagaacaaga gcgaaataaa agactggagc tgaaggaaca
     1801 ccaaccgtag gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     1861 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     1921 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     1981 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2041 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2101 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2161 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2221 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2281 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2341 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2401 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2461 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2521 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2581 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     2641 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     2701 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     2761 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     2821 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     2881 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     2941 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3001 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3061 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3121 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3181 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3241 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3301 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3361 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3421 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3481 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3541 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3601 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3661 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3721 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3781 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     3841 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     3901 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
     3961 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
     4021 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
     4081 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
     4141 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
     4201 gat
//
