LOCUS       sequence-413263-        3564 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 3564)
  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 3564)
  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-kcnk1b
FEATURES             Location/Qualifiers
     source          1..3564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     96..118
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     110..127
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     111..127
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    143..242
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1263..1362)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1379..1398)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1380..1398)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1403..1419)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1403..1419)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1521..1540)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1532..2341
                     /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(1599..1618)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2488..3076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2977..2996
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3230..3247
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3406..3433)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(join(3525..3564,1..47))
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
       61 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac
      121 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc
      181 gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagcagg
      241 cttcaccatg cttcagtccc tcgccggtaa ttcgtgcgtg cgcctgatac agagcaacaa
      301 atcggcgtgg tattttcttt tcctggtgct cggctacgtg ctgtatctga tcttcggtgc
      361 ggtggtgttt tcctcggtgg agctgccgta tgaagacgac cttcgccagc agctcagaga
      421 gataaagaga cttttcctcc aggagcacca gtgtctgtcc gaggagcggc tcgagaggtt
      481 tctgtccaaa gccctggagg ccagcaatta tggggtgtct attctcaaca acgcctcggc
      541 tagctggaac tgggacttca cctcgtcttt gttttttgcg agcaccgtct tatcgaccac
      601 aggatatggt cacactgccc ctctctctga tggcggaaag gcattctgca tcatttattc
      661 agtgattggc atccccttca ccctcctctt cctcacggct gtggtgcaaa ggatcatggt
      721 gtatagcacg tggaggccca tcatgtatgt tcacacacgc tggggtctgt ctaaacccct
      781 ggtggccctc gttcacgcga ctctgctggc tgtcgtggct gtgtcctgct tcttcctcat
      841 ccctgccgcc atcttctctg ccctagaaga gaactggaac tttctggagt ccttctactt
      901 ctgctttatc tccctttcca ccatcggcct tggtgactat gtgcccggag aggcctttaa
      961 ccagaggttt cgtgagctct acaaactggg catcactgtg tatctcctgc tgggcctgat
     1021 cgccatgctg gtggtgctgg agaccttctg tgagctccaa cagctcaagc agctccgcaa
     1081 gatgttttac ctgaagaagg agaaaccgca ggaccgtctg gccatcatgg agcacgacca
     1141 cctggccttc tcctccatgc cggacgccac catccactcg ccccacgagg acaaaaccga
     1201 gccctttgtg gacttccctg tgttgacctc tccaggagga gacgacccca tgatcaaata
     1261 ggacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     1321 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     1381 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     1441 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     1501 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     1561 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     1621 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     1681 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     1741 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     1801 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     1861 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     1921 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     1981 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2041 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2101 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2161 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     2221 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     2281 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     2341 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     2401 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     2461 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2521 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2581 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2641 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2701 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2761 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2821 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2881 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2941 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3001 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3061 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3121 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3181 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3241 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     3301 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     3361 aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc
     3421 aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac
     3481 cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg
     3541 agagcgttca ccgacaaaca acag
//
