LOCUS       sequence-413324-        3705 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 3705)
  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 3705)
  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-kcnk2b
FEATURES             Location/Qualifiers
     source          1..3705
                     /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(1314..1413)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1430..1449)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1431..1449)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1454..1470)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1454..1470)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1572..1591)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1583..2392
                     /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(1650..1669)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2539..3127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3028..3047
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3281..3298
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3457..3484)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3576..3662)
                     /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 cgctggaaga ccgtgctcag
      181 tgtcttcctg ctggtggtgc tttacctgat cctcggagca actgtgttta aggagctgga
      241 gcagccttat gagacgttac agaagctgaa catcctgatg gaaaaactgg agttccttga
      301 gcagcacccc tgcgtcaact catcagacct ggaaaactta gtcaagcagg tggtgtcagc
      361 attgcgggca ggagttaacc cttcaggaaa ctcgtccaat caaagcagtt tgtgggacct
      421 tagcaactcc ttcttcttct ctggaacggt tattacaact ataggatttg ggaatatctc
      481 gccacacact gaagtcggac gcatcttctg tattatttat gctctgctgg gaattcctct
      541 gttcggattc ttgctggcag gagtgggaga ccagctcgga accatctttg gaaaggcaat
      601 tgccaaagtg gagggtatga ttgataagtg gaacgtgagt caaaccaaga tccgggtgat
      661 ctctactctt cttttcattc tgtttggctg tctgctcttc gtcacgctgc cggccgttat
      721 attcaagcac atcgagggct ggagcgcact ggagtccatc tactttgtcg tcattacctt
      781 gacaaccatt ggttttggag atttcgttgc aggcgaggct gagcgacgcc accatgaaga
      841 gagcagtgga ggttctcagc tggagtacct ggactactac aagcccctgg tgtggttttg
      901 gatcctggtt ggtttagcct acttcgctgc tgttctcagt atgattggag actggttccg
      961 ggttatatcc aagaaaacaa aagaagaggt aggagagttt cgagctcatg ctgccgaatg
     1021 gacggctaat gtgtccgctg agtttaaaga gacacgtcgt cgactgagtg tcgacattta
     1081 tgacaaattc cagcgtgcaa cttcagtaaa acgcaaactg tctgcggaaa taaatctcag
     1141 ccctcctatc aatcagcaga tgaccccagg gaagcgtgca cgttcggtta accttggaga
     1201 cgagagagag gcatatccct acacattagc acgaaacgga agtctattcc tcaacagcct
     1261 catcccggat tacgctgacc atcgggatat gactagaata cagacaaaat gagacccagc
     1321 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     1381 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     1441 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     1501 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1561 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     1621 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     1681 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1741 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1801 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     1861 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     1921 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     1981 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     2041 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2101 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     2161 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2221 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2281 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2341 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2401 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     2461 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     2521 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2581 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2641 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2701 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2761 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2821 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2881 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2941 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3001 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3061 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3121 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3181 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3241 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3301 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3361 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac
     3421 gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
     3481 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
     3541 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     3601 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
     3661 tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggat
//
