LOCUS       sequence-413244-        3831 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 3831)
  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 3831)
  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-kcnk2a
FEATURES             Location/Qualifiers
     source          1..3831
                     /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(1440..1539)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1556..1575)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1557..1575)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1580..1596)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1580..1596)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1698..1717)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1709..2518
                     /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(1776..1795)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2665..3253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3154..3173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3407..3424
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3583..3610)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3702..3788)
                     /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 aggagcagcg agaacagcag
      181 atcccacaaa gtagtaatgg ctgcacctga tcttttagat cctaaatcag ccacgcacaa
      241 caccaaacca cggctgtcat tctcctccaa gcccatcgtg tacaactctg gagatgattg
      301 tgagagcata accaccgtaa tgaagtggaa aacagtgctg gccatcttcc tgctggtggt
      361 cctgtacctg atcatcggtg ccactgtgtt caaagcactg gaacagcctg aggagggctt
      421 gcagaagtac cgcatcatcc aggagaagat cgactttctc tcaatgcaca cctgtgtcaa
      481 cacctcggag ctggaggatc tggtcaagca agtggtgtta gccatccggg ccggggtcaa
      541 tccatccggt cacccctcca atgagagcag catgtgggac ctcagcagct ccttcttctt
      601 tgctgggacc gtcatcacta ctatagggtt tggaaatgtt tctccacaca cagaaggagg
      661 caggatcttc tgcattatct acgccctgtt ggggatccca ctctttggct ttctcttggc
      721 tggagtgggc gatcagctgg gaaccatatt tggcaaagga atcgccaaag tggaaaagat
      781 gtttgtgaaa tggaacgtga gccagactaa aatccgtgtg acgtccacag tcctcttcat
      841 cctgttcggg tgtcttctgt ttgtggctct tcctgccctt atcttccagc acattgaagg
      901 ctggagcgct ctcgagtcca tctactttgt tgtcatcact ttgaccacaa ttggattcgg
      961 agactttgtg gcagggggct cggagattga atatctggac tattataagc ccatcgtatg
     1021 gttctggatc ctggtgggtc ttgcgtactt tgctgctgtt ctcagtatga ttggagactg
     1081 gcttcgggtc atctccaaaa agaccaaaga ggaggtggga gagtttcgtg ctcacgctgc
     1141 agaatggacg gctaacgtgt ccgctgagtt taaagagacg cgccgacgcc tgagcgtgga
     1201 aatctatgac aagtttcagc gggccgcgta catcaaacgc aagctgtcat cagagctggg
     1261 ccaaaaccca ggccaggaca tgatgccatg caagcgaacc ctgtctgtaa acttcacaga
     1321 tgaactggag aaggaaggtc tccctacttt gaccaaaaac ggcagtctct atctgaatgg
     1381 gcttactcca gactgtcccg attgcagtga gatttccatc attgaacatc tcaaataaga
     1441 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     1501 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     1561 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     1621 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     1681 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     1741 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     1801 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     1861 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     1921 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     1981 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     2041 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2101 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2161 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     2221 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     2281 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     2341 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     2401 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     2461 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     2521 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     2581 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     2641 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2701 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2761 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2821 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2881 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2941 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3001 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3061 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3121 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3181 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3241 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3301 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3361 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3421 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3481 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3541 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     3601 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
     3661 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
     3721 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt
     3781 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga t
//
