LOCUS       sequence-413351-        3897 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 3897)
  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 3897)
  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-kcnk5b
FEATURES             Location/Qualifiers
     source          1..3897
                     /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(1506..1605)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1622..1641)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1623..1641)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1646..1662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1646..1662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1764..1783)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1775..2584
                     /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(1842..1861)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2731..3319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3220..3239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3473..3490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3649..3676)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3768..3854)
                     /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 gcagataaag gacctatttt
      181 gacatcagtc ataattttct acttgtccat tggagctgcg atattccaaa tcctcgagga
      241 gccgaattta aattcagctg tggacgatta taaaaataaa actaacaatt tattgaagaa
      301 atatccctgc ctaagtaaag aggtcttggg cgaaattatt gaggtagtgg cggaggcgac
      361 tggacaaggt gtgactgtaa caaaagaagt acaattcaac aattggaatt gggagaatgc
      421 tgttattttt gctgctacag tcatcactac tataggttat ggtaatgttg ctccaaagac
      481 aactggaggt cgtttattct gtatcctcta tggtctctgt ggcattcctc tgtgcctcac
      541 ctggataagt gagctgggta cattttttgg cagcaggaca aagcgtctaa gtcagttgct
      601 cctccacagt ggcctcaatg tgagaaaagt tcagttcatc tgcacaattg tgttcctttt
      661 atggggcttc ttggttcact taatcatccc agcctttgtg ttcatgttct ttgagaattg
      721 gacttactta gaaggcctgt acttctcctt caccaccctg acgactgtcg gttttggtga
      781 ctatgtggca ggggtggacc catccgtaaa ctaccccact ctctacagat tctttgtcca
      841 gctatggatc tacctgggcc ttgcttggtt gtcattgttc ttcagttgga acgtgcatat
      901 ggtggtggaa gctcataaag ttttaaagaa aaggaggatg agacgccaca ggctgcccac
      961 cgatgacgtg ccggaaaaaa aagaagtgaa aaaaacacca aagccacctc ctcgctctgg
     1021 agtcattgac atctttgagt tcatgtccga gaaggtcgag gactacagcg acgttatccg
     1081 agctatcgga gcagacgaaa agagaaggaa aaagaaacag gaggaggaac tggcacgatc
     1141 caagagctgc agtgaccttt tgcaaggcct ggtcatcccg cttgatcact ctcctcgtct
     1201 gcaacggcgg ttcagtgtca gcgccaacat gtgcatggcc atatctgatg agtctgtgga
     1261 tggcctcaac aataacaatt gcaaacaaga ggatgacaca ctaactaaag tgcgccataa
     1321 agaccagaag gttaatcgag agagagcgga gaatcctgca cgttgtgcat gggactctag
     1381 atcatctgat ccatcaatct ttcaaagttc cactgttacc aacagcacta atagaggatc
     1441 aagattctca gtgtccaaag tttcagaaga tcgtttatta gggaagagga aaagctttgg
     1501 ttgagaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     1561 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     1621 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1681 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1741 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     1801 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1861 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     1921 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1981 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2041 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2101 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2161 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2221 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2281 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2341 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2401 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2461 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2521 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2581 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     2641 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     2701 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2761 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2821 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2881 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2941 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3001 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3061 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3421 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3481 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     3541 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     3601 cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc
     3661 gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc
     3721 caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc
     3781 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc
     3841 tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggat
//
